PK!gK0%%celltk/__init__.pyfrom labeledarray import LabeledArrayPK!P##celltk/apply.py""" python celltk/apply.py -i c0/img_00000000* -l c2/img_00000000* TODO: need to deal with parent id """ from utils.util import imread import argparse import tifffile as tiff from os.path import basename, join, dirname, abspath import numpy as np from utils.postprocess_utils import regionprops, Cell # set default parent and next as None try: from labeledarray import LabeledArray except: from celltk.labeledarray import LabeledArray from os.path import exists from utils.file_io import make_dirs, lbread import pandas as pd import logging from scipy.ndimage.morphology import binary_fill_holes from scipy.ndimage.morphology import binary_dilation import warnings warnings.filterwarnings('ignore', category=np.VisibleDeprecationWarning) warnings.filterwarnings('ignore', category=pd.io.pytables.PerformanceWarning) logger = logging.getLogger(__name__) PROP_SAVE = ['area', 'cell_id', 'convex_area', 'cv_intensity', 'eccentricity', 'major_axis_length', 'minor_axis_length', 'max_intensity', 'mean_intensity', 'median_intensity', 'min_intensity', 'orientation', 'perimeter', 'solidity', 'std_intensity', 'total_intensity', 'x', 'y', 'parent', 'num_seg'] MAX_NUMCELL = 100000 def find_all_children(labels): mask = binary_fill_holes(labels < 0) mask[labels < 0] = False clabelnums = np.unique(labels[mask]).tolist() if 0 in clabelnums: clabelnums.remove(0) return clabelnums def find_parent_label(labels, child_label): mask = binary_dilation(labels == child_label) mask[labels == child_label] = False return max(set(labels[mask].tolist()), key=labels[mask].tolist().count) def add_parent(cells, labels): children_labels = find_all_children(labels) for cl in children_labels: parent_label = find_parent_label(labels, cl) child = [cell for cell in cells if cell.label == cl] assert len(child) == 1 child[0].parent = abs(parent_label) return cells def apply(): pass def initialize_arr(store): nframe = len(store) cell_ids = [[i.cell_id for i in cells] for cells in store] cell_ids = [i for j in cell_ids for i in j] ncells = len(np.unique(cell_ids)) return np.zeros((len(PROP_SAVE), ncells, nframe)), np.unique(cell_ids).tolist() def df2larr(df): index = df.index.tolist() frames = len(np.unique([i[-1] for i in index])) index_without_frame = list(set([i[:-1] for i in index])) arr = np.empty((df.shape[0]/frames, df.shape[1], frames)) arr[:] = np.nan arr_labels = [] for num, ind in enumerate(index_without_frame): arr_labels.append(ind) arr[num, :, :] = df.ix[ind].values.T larr = LabeledArray(arr, arr_labels) larr.time = np.arange(arr.shape[-1]) return larr def _cells2array(cells): arr = np.zeros((len(cells), len(PROP_SAVE)), np.float32) for cnum, cell in enumerate(cells): arr[cnum, :] = [getattr(cell, k) for k in PROP_SAVE] return arr # def multi_index(cells, obj_name, ch_name): # frames = np.unique([i.frame for i in cells]) # index = pd.MultiIndex.from_product([obj_name, ch_name, PROP_SAVE, frames], names=['object', 'ch', 'prop', 'frame']) # column_idx = pd.MultiIndex.from_product([np.unique([i.cell_id for i in cells])]) # df = pd.DataFrame(index=index, columns=column_idx, dtype=np.float32) # for cell in cells: # for k in PROP_SAVE: # df[cell.cell_id].loc[obj_name, ch_name, k, cell.frame] = np.float32(getattr(cell, k)) # return df def caller(inputs_list, inputs_labels_list, output, primary, secondary): make_dirs(dirname(abspath(output))) inputs_list = [inputs_list, ] if isinstance(inputs_list[0], str) else inputs_list inputs_labels_list = [inputs_labels_list, ] if isinstance(inputs_labels_list[0], str) else inputs_labels_list obj_names = [basename(dirname(i[0])) for i in inputs_labels_list] if primary is None else primary ch_names = [basename(dirname(i[0])) for i in inputs_list] if secondary is None else secondary for inputs, ch in zip(inputs_list, ch_names): for inputs_labels, obj in zip(inputs_labels_list, obj_names): logger.info("Channel {0}: {1} applied...".format(ch, obj)) arr = np.ones((MAX_NUMCELL, len(PROP_SAVE), len(inputs)), np.float32) * np.nan for frame, (path, pathl) in enumerate(zip(inputs, inputs_labels)): img, labels = imread(path), lbread(pathl, nonneg=False) cells = regionprops(labels, img) if (labels < 0).any(): cells = add_parent(cells, labels) [setattr(cell, 'frame', frame) for cell in cells] cells = [Cell(cell) for cell in cells] tarr = _cells2array(cells) index = tarr[:, 1].astype(np.int32) arr[index, :, frame] = tarr logger.info("\tmaking dataframe...") cellids = np.where(~np.isnan(arr[:, 0, :]).all(axis=1))[0] marr = np.zeros((len(cellids), arr.shape[1], arr.shape[2])) for pn, i in enumerate(cellids): marr[pn] = arr[i] sarr = np.swapaxes(marr, 0, 2) narr = sarr.reshape((sarr.shape[0]*sarr.shape[1], sarr.shape[2]), order='F') index = pd.MultiIndex.from_product([obj, ch, PROP_SAVE, range(arr.shape[-1])], names=['object', 'ch', 'prop', 'frame']) df = pd.DataFrame(narr, index=index, columns=cellids) if exists(join(output, 'df.csv')): ex_df = pd.read_csv(join(output, 'df.csv'), index_col=['object', 'ch', 'prop', 'frame']) ex_df.columns = pd.to_numeric(ex_df.columns) ex_df = ex_df.astype(np.float32) df = pd.concat([df, ex_df]) df.to_csv(join(output, 'df.csv')) larr = df2larr(df) larr.save(join(output, 'df.npz')) logger.info("\tdf.npz saved.") def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--inputs", help="images", nargs="*", default=[]) parser.add_argument("-l", "--labels", help="labels", nargs="+") parser.add_argument("-o", "--output", help="file name", type=str, default='temp') parser.add_argument("-p", "--primary", help="name for primary key, typically objects", type=str, default=None) parser.add_argument("-s", "--secondary", help="name for secondary key, typically channels", type=str, default=None) args = parser.parse_args() output = args.output caller(args.inputs, args.labels, args.output, args.primary, args.secondary) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!''celltk/caller.py import argparse from os.path import join, isdir, exists from glob import glob import logging from logging import FileHandler, StreamHandler import yaml import multiprocessing from utils.file_io import make_dirs import sys logger = logging.getLogger(__name__) import os from os.path import join, basename, exists, dirname import collections def multi_call(inputs): contents = load_yaml(inputs) pin = contents['PARENT_INPUT'] pin = pin[:-1] if pin.endswith('/') or pin.endswith('\\') else pin input_dirs = [join(pin, i) for i in os.listdir(pin) if os.path.isdir(join(pin, i))] contents_list = [] for subfolder in input_dirs: conts = eval(str(contents).replace('$INPUT', subfolder)) conts['OUTPUT_DIR'] = join(conts['OUTPUT_DIR'], basename(subfolder)) contents_list.append(conts) return contents_list def convert(data): if isinstance(data, basestring): return str(data) elif isinstance(data, collections.Mapping): return dict(map(convert, data.iteritems())) elif isinstance(data, collections.Iterable): return type(data)(map(convert, data)) else: return data def extract_path(path): f = glob(path) if isdir(f[0]) or not f: f = glob(join(path, '*')) return f def parse_lazy_syntax(inputs, outputdir): if isinstance(inputs, str): in0 = sorted(glob(inputs)) if not in0: in0 = sorted(glob(join(outputdir, inputs))) if isdir(in0[0]): in0 = sorted(glob(join(in0[0], '*'))) elif isinstance(inputs, list): if all([exists(i) for i in inputs]): return inputs in0 = zip(*[sorted(glob(i)) for i in inputs]) if not in0: in0 = zip(*[sorted(glob(join(i, '*'))) for i in inputs]) if not in0: in0 = zip(*[sorted(extract_path(join(outputdir, i))) for i in inputs]) return in0 def prepare_path_list(inputs, outputdir): try: in0 = parse_lazy_syntax(inputs, outputdir) except IndexError: logger.info("Images \"{0}\" not found. Check your path".format(inputs)) print "Images \"{0}\" not found. Check your path".format(inputs) sys.exit(1) return in0 def retrieve_in_list(obj, key, empty=[]): if isinstance(obj, dict): obj = [obj, ] st = [] for ob in obj: if key not in ob: st.append(empty) else: st.append(ob[key]) return st def parse_operation(operation): functions = retrieve_in_list(operation, 'function') params = retrieve_in_list(operation, 'params', empty={}) images = retrieve_in_list(operation, 'images')[0] labels = retrieve_in_list(operation, 'labels')[0] output = retrieve_in_list(operation, 'output')[-1] return functions, params, images, labels, output def _retrieve_caller_based_on_function(function): import preprocess, segment, track, postprocess, subdetect, apply import preprocess_operation, segment_operation, track_operation, postprocess_operation, subdetect_operation ops_modules = [preprocess_operation, segment_operation, track_operation, postprocess_operation, subdetect_operation, apply] caller_modules = [preprocess, segment, track, postprocess, subdetect, apply] module = [m for m, top in zip(caller_modules, ops_modules) if hasattr(top, function)][0] return getattr(module, "caller") def run_operation(output_dir, operation): functions, params, images, labels, output = parse_operation(operation) inputs = prepare_path_list(images, output_dir) logger.info(inputs) inputs_labels = prepare_path_list(labels, output_dir) output = join(output_dir, output) if output else output_dir caller = _retrieve_caller_based_on_function(functions[0]) if len(functions) == 1 and functions[0] == 'apply': ch_names = operation['ch_names'] if 'ch_names' in operation else images obj_names = operation['obj_names'] if 'obj_names' in operation else labels caller(zip(*inputs), zip(*inputs_labels), output, obj_names, ch_names) elif not inputs_labels: caller(inputs, output, functions, params=params) else: caller(inputs, inputs_labels, output, functions, params=params) def run_operations(output_dir, operations): for operation in operations: run_operation(output_dir, operation) def load_yaml(path): with open(path) as stream: contents = yaml.load(stream) return contents def single_call(inputs): contents = load_yaml(inputs) call_operations(contents) def call_operations(contents): make_dirs(contents['OUTPUT_DIR']) logging.basicConfig(filename=join(contents['OUTPUT_DIR'], 'log.txt'), level=logging.DEBUG) logging.getLogger("PIL").setLevel(logging.WARNING) run_operations(contents['OUTPUT_DIR'], contents['operations']) logger.info("Caller finished.") return def _parallel(args): ''' Use this function if you want to multiprocess using PARENT_INPUT argument (see input_fireworks.yml). ''' contents_list = multi_call(args.input[0]) contents_list = [convert(i) for i in contents_list] pool = multiprocessing.Pool(args.cores, maxtasksperchild=1) pool.map(call_operations, contents_list, chunksize=1) pool.close() def parse_args(): parser = argparse.ArgumentParser() parser.add_argument("-n", "--cores", help="number of cores for multiprocessing", type=int, default=1) parser.add_argument("input", nargs="*", help="input argument file path") args = parser.parse_args() return args def main(): args = parse_args() if len(args.input) == 1: contents = load_yaml(args.input[0]) if "PARENT_INPUT" in contents: _parallel(args) else: call_operations(contents) # single_call(args.input[0]) if len(args.input) > 1: num_cores = args.cores print str(num_cores) + ' started parallel' pool = multiprocessing.Pool(num_cores, maxtasksperchild=1) pool.map(single_call, args.input, chunksize=1) pool.close() if __name__ == "__main__": main() PK!Cjcelltk/caller_parallel.py''' Call this function if you want to multiprocess using PARENT_INPUT argument (see input_fireworks.yml). If you simply want to pass multiple input files, use caller.py. ''' from __future__ import print_function import os import sys from os.path import join, basename, exists, dirname celltkroot = dirname(dirname(os.path.abspath(__file__))) sys.path.append(join(celltkroot, 'celltk')) # sys.path.insert(0, join(celltkroot, 'celltk')) from caller import parse_args, load_yaml, call_operations import yaml import collections import multiprocessing def convert(data): if isinstance(data, basestring): return str(data) elif isinstance(data, collections.Mapping): return dict(map(convert, data.iteritems())) elif isinstance(data, collections.Iterable): return type(data)(map(convert, data)) else: return data def parallel_call(contents): contents = convert(contents) call_operations(contents) def multi_call(inputs): contents = load_yaml(inputs) pin = contents['PARENT_INPUT'] pin = pin[:-1] if pin.endswith('/') or pin.endswith('\\') else pin input_dirs = [join(pin, i) for i in os.listdir(pin)] contents_list = [] for subfolder in input_dirs: conts = eval(str(contents).replace('$INPUT', subfolder)) conts['OUTPUT_DIR'] = join(conts['OUTPUT_DIR'], basename(subfolder)) contents_list.append(conts) return contents_list def main(): args = parse_args() contents_list = multi_call(args.input[0]) num_cores = args.cores print(str(num_cores) + ' started parallel') pool = multiprocessing.Pool(num_cores, maxtasksperchild=1) pool.map(parallel_call, contents_list, chunksize=1) pool.close() if __name__ == "__main__": main() PK!K""celltk/command.pyfrom caller import _retrieve_caller_based_on_function from utils.parser import ParamParser, parse_image_files import argparse from utils.global_holder import holder from utils.file_io import make_dirs, imsave from utils.util import imread def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--inputs", help="images", nargs="*") parser.add_argument("-l", "--inputs_labels", help="images", nargs="*", default=None) parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="*") parser.add_argument("-p", "--param", nargs="+", help="parameters", action='append') args = parser.parse_args() params = ParamParser(args.param).run() inputs = parse_image_files(args.inputs) if args.inputs_labels is not None: args.inputs_labels = parse_image_files(args.inputs_labels) caller = _retrieve_caller_based_on_function(args.functions[0]) if len(args.functions) == 1 and args.functions[0] == 'apply': pass # ch_names = operation['ch_names'] if 'ch_names' in operation else images # obj_names = operation['obj_names'] if 'obj_names' in operation else labels # caller(zip(*inputs), zip(*args.inputs_labels), args.output, obj_names, ch_names) elif args.inputs_labels is None: caller(inputs, args.output, args.functions, params=params) else: caller(inputs, args.inputs_labels, args.output, args.functions, params=params) if __name__ == "__main__": main() PK! *((celltk/labeledarray/LICENSEMIT License Copyright (c) 2017 braysia Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions: The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software. THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE. PK!celltk/labeledarray/README.md# LabeledArray Numpy array subclass for indexing by strings. Using multi-index in pandas sometimes provides complications in terms of "copies vs views". This array is to provide numpy.array's behavior and still enable to slice array by strings. Underlying data can be 2D, 3D or N-dimensional array. First dimension will be used for labels (multi-index). ``` arr = np.zeros((3, 20, 100)) labels = np.array([['nuc' ,'area', ''], ['nuc' ,'FITC' , 'min_intensity'], ['nuc' ,'FITC' , 'max_intensity']], dtype=object) larr = LabeledArray(arr, labels) print larr.shape print larr['nuc', 'FITC'].shape print larr['nuc', 'FITC', 'max_intensity'].shape ``` The extra attributes including labels are automatically saved and loaded with the array. ``` larr = LabeledArray(arr, labels) larr.time = np.arange(arr.shape[-1]) larr.save('temp') new_larr = LabeledArray().load('temp') print new_larr.time ``` PK!ي22celltk/labeledarray/__init__.pyfrom labeledarray.labeledarray import LabeledArrayPK!,celltk/labeledarray/labeledarray/__init__.pyPK!qq0celltk/labeledarray/labeledarray/labeledarray.py""" TODO: check if labels is unique. """ import numpy as np from collections import OrderedDict from utils import sort_labels_and_arr, uniform_list_length class LabeledArray(np.ndarray): """ Each rows corresponds to labels, each columns corresponds to cells. Underlying data structure can be N-dimensional array. First dimension will be used for labeled array. Examples: >> arr = np.arange(12).reshape((3, 2, 2)) >> labelarr = np.array([['a1' ,'b1', ''], ['a1' ,'b2' , 'c1'], ['a1' ,'b2' , 'c2']], dtype=object) >> darr = DArray(arr, labelarr) >> assert darr['a1'].shape (3, 2, 2) >> darr['a1', 'b1'].shape (2, 2) >> darr['a1', 'b2', 'c1'] DArray([[4, 5], [6, 7]]) """ idx = None labels = None def __new__(cls, arr=None, labels=None, idx=None): if arr is None: return np.asarray(arr).view(cls) labels, arr = sort_labels_and_arr(labels, arr) if not isinstance(labels, np.ndarray) and labels is not None: labels = np.array(uniform_list_length(labels), dtype=object) obj = np.asarray(arr).view(cls) obj.labels = labels obj.idx = idx return obj def __array_finalize__(self, obj): if obj is None: return self.labels = getattr(obj, 'labels', None) if self.labels is None: return if hasattr(obj, 'idx') and self.ndim >= 1: if obj.idx is None: return if isinstance(obj.idx, int): self.labels = self.labels[obj.idx] else: self.labels = self.labels[obj.idx[0]] if isinstance(self.labels, str): return if self.labels.ndim > 1: f_leftshift = lambda a1:all(x>=y for x, y in zip(a1, a1[1:])) all_column = np.all(self.labels == self.labels[0,:], axis=0) sl = 0 if not f_leftshift(all_column) else all_column.sum() self.labels = self.labels[:, slice(sl, None)] if self.labels.ndim == 1: self.labels = None obj = np.array(obj) def __getitem__(self, item): if isinstance(item, str): item = self._label2idx(item) if isinstance(item, tuple): if isinstance(item[0], str): item = self._label2idx(item) self.idx = item ret = super(LabeledArray, self).__getitem__(item) return ret.squeeze() def _label2idx(self, item): item = (item, ) if not isinstance(item, tuple) else item boolarr = np.ones(self.labels.shape[0], dtype=bool) for num, it in enumerate(item): boolarr = boolarr * (self.labels[:, num]==it) tidx = np.where(boolarr)[0] if boolarr.sum() == 1: return tuple(tidx) if boolarr.all(): return (slice(None, None, None), ) + (slice(None, None, None), ) * (self.ndim - 1) minidx = min(tidx) if min(tidx) > 0 else None maxidx = max(tidx)+1 if max(tidx)+1 < self.shape[0] else None if boolarr.sum() > 1: return (slice(minidx, maxidx, None), ) + (slice(None, None, None), ) * (self.ndim - 1) def vstack(self, larr): """merging first dimension (more labels) """ if self.ndim > larr.ndim: larr = np.expand_dims(larr, axis=0) return LabeledArray(np.vstack((self, larr)), np.vstack((self.labels, larr.labels))) def hstack(self, larr): """merging second dimension (more cells) """ if (self.labels == larr.labels).all(): return LabeledArray(np.hstack((self, larr)), self.labels) def save(self, file_name): extra_fields = set(dir(self)).difference(set(dir(self.__class__))) data = dict(arr=self, labels=self.labels) for ef in extra_fields: data[ef] = getattr(self, ef) np.savez_compressed(file_name, **data) @classmethod def load(cls, file_name): if not file_name.endswith('.npz'): file_name = file_name + '.npz' f = np.load(file_name) arr, labels = f['arr'], f['labels'] la = LabeledArray(arr, labels) for key, value in f.iteritems(): if not ('arr' == key or 'labels' == key): setattr(la, key, value) return la if __name__ == "__main__": # Check 2D. arr = np.random.rand(3, 100) labelarr = np.array([['a1', 'b1', ''], ['a1' ,'b2' , 'c1'], ['a1' ,'b2' , 'c2']], dtype=object) darr = LabeledArray(arr, labelarr) # stop assert darr['a1'].shape == (3, 100) assert darr['a1', 'b1'].shape == (100, ) assert darr['a1', 'b2'].shape == (2, 100) assert darr['a1', 'b2', 'c1'].shape == (100, ) # check 3D. arr = np.arange(12).reshape((3, 2, 2)) labelarr = np.array([['a1' ,'b1', ''], ['a1' ,'b2' , 'c1'], ['a1' ,'b2' , 'c2']], dtype=object) darr = LabeledArray(arr, labelarr) assert darr['a1'].shape == (3, 2, 2) assert darr['a1', 'b1'].shape == (2, 2) assert darr['a1', 'b2'].shape == (2, 2, 2) assert darr['a1', 'b2', 'c1'].shape == (2, 2) assert darr.shape == (3, 2, 2) assert darr[1:, :, :].shape == (2, 2, 2) assert darr[1, :, :].shape == (2, 2) assert np.all(darr['a1', 'b2'].labels == np.array([['c1'], ['c2']])) # can save and load extra fields. add "time" for example. darr.time = np.arange(darr.shape[-1]) darr.save('test') cc = LabeledArray().load('test.npz') assert cc.time.shape == (2,) cc[0:2, :, :] cc['a1', 'b1'][0, 0] = 100 assert np.sum(cc == 100) == 1 assert darr.vstack(darr).shape == (2 * darr.shape[0], darr.shape[1], darr.shape[2]) assert darr.hstack(darr).shape == (darr.shape[0], 2 * darr.shape[1], darr.shape[2]) PK!A2G)celltk/labeledarray/labeledarray/temp.npzPK"uJ&PParr.npyNUMPYF{'descr': '+U@1U@U@ NT@wT@ҔT@`T@@hT@ T@ T@ S@S@T@T@ NU@@U@ ,U@T@T@YT@9U@,OU@`fU@ PU@hU@_U@@OU@@)*U@T@@T@@ T@8k@-xj@ "j@@oi@i@@j@@j@,3k@5k@Vk@:mk@k@"k@k@k@k@ Zk@!k@ k@4k@ k@Xk@sj@j@zj@Hj@Wj@@wcj@tj@+{j@`jj@`\j@[j@ +>j@Lj@j@`i@i@Hi@ x i@MFp@p@`'o@ Ho@\mn@m@m@@l@_k@ k@\j@j@j@j@mij@`Nj@`?4j@) j@Wi@`?4i@`z#i@mh@ h@h@F`h@$Fh@>h@h@h@ Ih@g@Bg@g@mg@h@Ph@@h@g@g@g@`y@b%z@ sz@z@[{@@z@z@z@@z@z@`pz@ |Xz@@Cz@z@`_z@oy@ظy@jx@ x@ *v@ɸu@u@u@u@@u@-v@@v@v@@i>v@ 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V?e?u?@I???2N? z??Z??@?{?c? ;?@۬?O6?@6??ڿ??@H?P?? ?`%Q?@T?@T?z?k7? qX?L?H?? ? u?@ ?? ?Ϩ?@S? ?? w?s?5?P?@9?@'?Q(?(?`;? ?8?&? f?`2?@??`?@ ?^`??`?1?%?@c\?a??9?PK"uJ.arr_labels.npyNUMPYF{'descr': '|S17', 'fortran_order': False, 'shape': (72, 3), } cytoDAPIxnucDAPIcell_idnucDAPIxcytoYFPmedian_intensitycytoYFPareanucDAPIperimeternucYFPareacytoDAPIperimetercytoDAPImin_intensitynucYFPynucYFPcell_idcytoYFPtotal_intensitynucYFPperimetercytoYFPmajor_axis_lengthnucDAPImajor_axis_lengthnucDAPImedian_intensitycytoYFPorientationcytoYFPstd_intensitynucYFPminor_axis_lengthnucYFPsoliditycytoDAPIareanucDAPIstd_intensitynucDAPIsoliditycytoDAPIstd_intensitycytoYFPconvex_areanucYFPmean_intensitynucDAPItotal_intensitynucYFPmedian_intensitynucYFPtotal_intensitynucDAPImax_intensitynucDAPIeccentricitycytoDAPIcv_intensitycytoDAPIycytoDAPImean_intensitynucYFPxcytoDAPIorientationcytoYFPperimetercytoYFPminor_axis_lengthnucDAPImean_intensitynucYFPmax_intensitycytoYFPsoliditynucDAPIareacytoDAPIsoliditynucDAPIorientationnucDAPImin_intensitycytoYFPcell_idcytoYFPycytoDAPIcell_idnucDAPIycytoDAPIeccentricitynucYFPmajor_axis_lengthcytoYFPxcytoDAPIconvex_areacytoYFPmin_intensitycytoDAPItotal_intensitynucDAPIminor_axis_lengthcytoYFPcv_intensitynucYFPorientationcytoYFPmax_intensitynucYFPconvex_areanucYFPmin_intensitynucYFPeccentricitycytoDAPImax_intensitycytoYFPeccentricitynucDAPIcv_intensitycytoDAPImedian_intensitynucDAPIconvex_areacytoYFPmean_intensitynucYFPstd_intensitycytoDAPImajor_axis_lengthcytoDAPIminor_axis_lengthnucYFPcv_intensityPK"uJ&PParr.npyPK"uJ.uarr_labels.npyPKqIPK!y+*)celltk/labeledarray/labeledarray/test.npzPKuJefarr.npyPZ\nni_TR_ wK)Ng$: F@P\ P43fҬP JCi( arCiPKuJFi labels.npyEOK0ƓT̾DnP . v2iQakM xk {ϟO_(7Ƣ|򰔑 =rSaynv0{8."~dlu ]ow o٪l>> labels = [['a', 'B', '1'], ['a', 'A', '1'], ['b', 'A', '3'], ['b', 'B', '2']] >>> sort_labels_and_arr(labels) [['a', 'A', '1'], ['a', 'B', '1'], ['b', 'A', '3'], ['b', 'B', '2']] >>> labels = [['a', 'B', '1'], ['prop'], ['aprop'], ['b', 'B', '2']] >>> sort_labels_and_arr(labels) [['a', 'B', '1'], ['aprop'], ['b', 'B', '2'], ['prop']] ''' labels = [list(i) for i in labels] labels, sort_idx = sort_multi_lists(labels) if not len(arr): return labels if len(arr): arr = arr[sort_idx] return labels, arr def uniform_list_length(labels): """ Insert empty string untill all the elements in labels have the same length. Examples: >>> uniform_list_length([['a'], ['a', 'b'], ['a', 'b', 'c']]) [['a', ' ', ' '], ['a', 'b', ' '], ['a', 'b', 'c']] """ max_num = max([len(i) for i in labels]) for label in labels: for num in range(1, max_num): if len(label) == num: label.extend([" " for i in range(max_num - num)]) return labels def undo_uniform_list_length(labels): """ Remove empty string after the operation done by uniform_list_length. Examples: >>> undo_uniform_list_length(uniform_list_length([['a'], ['a', 'b'], ['a', 'b', 'c']])) [['a'], ['a', 'b'], ['a', 'b', 'c']] """ for label in labels: while " " in label: label.remove(" ") return labels def sort_multi_lists(labels): """ Sort a list by the order of column 0, 1 and 2.... Works for a list having different length of elements. Examples: >>> sort_multi_lists([['a', 'c'], ['a', 'b'], ['a', 'b', 'c']]) ([['a', 'b'], ['a', 'b', 'c'], ['a', 'c']], [1, 2, 0]) """ unilabels = uniform_list_length(labels) intlist = [[i] * 3 for i in range(len(unilabels))] sort_func = lambda item: [i for i in item[0]] sort_idx = [ii[0] for (i, ii) in sorted(zip(unilabels, intlist), key=sort_func)] sort_labels = [unilabels[i] for i in sort_idx] return undo_uniform_list_length(sort_labels), sort_idx PK!5QPPcelltk/postprocess.py""" python celltk/postprocess.py -f gap_closing -i c0/img_00000000* -l c2/img_00000000* -o c3 -p DISPLACEMENT=30 """ from scipy.ndimage import imread import argparse import tifffile as tiff from os.path import basename, join import numpy as np import postprocess_operation from utils.postprocess_utils import regionprops, LCell # set default parent and next as None from utils.file_io import make_dirs, imsave, lbread from utils.filters import labels2outlines from scipy.ndimage import binary_dilation from utils.parser import ParamParser from utils.global_holder import holder import logging logger = logging.getLogger(__name__) def neg2poslabels(labels): maxint = labels.max() negatives = np.unique(labels[labels < 0]) for i in negatives: maxint += 1 labels[labels == i] = maxint return labels def cells2labels0(cells, frame, shape): inframe = [i for i in cells if i.frame == frame] template = np.zeros(shape) for cell in inframe: for c0, c1 in cell.coords: template[c0, c1] = cell.label return template def cells2labels(cells, frame, labels): inframe = [i for i in cells if i.frame == frame] template = np.zeros(labels.shape) for cell in inframe: template[labels == cell._original_label] = cell.label if cell.parent is not None: outline = labels2outlines(binary_dilation(template == cell.label)) # outline = labels2outlines(template == cell.label) template[outline > 0] = -cell.parent return template def caller(inputs, inputs_labels, output, functions, params): make_dirs(output) # Make cells. cells are a list of regionproperties or subclasses. logger.info('Postprocess.\tcollecting cells...') store = [] for frame, (path, pathl) in enumerate(zip(inputs, inputs_labels)): img, labels = imread(path), lbread(pathl) cells = regionprops(labels, img) cells = [LCell(cell) for cell in cells] for cell in cells: cell.frame = frame if frame > 0: all_labels = [i.label for i in store[frame - 1]] if cell.label in all_labels: store[frame - 1][all_labels.index(cell.label)].nxt = cell store.append(cells) cells = [i for j in store for i in j] # Each function receives cells (regionprops) and finally return labels generated by cells.label for function, param in zip(functions, params): logger.info('\trunning {0}'.format(function)) func = getattr(postprocess_operation, function) cells = func(cells, **param) logger.info('\tsaving images...') for frame, (path, pathl) in enumerate(zip(inputs, inputs_labels)): labels = cells2labels(cells, frame, lbread(pathl)) imsave(labels, output, path, dtype=np.int16) def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--input", help="images", nargs="+") parser.add_argument("-l", "--labels", help="labels", nargs="+") parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="+") parser.add_argument('-p', '--param', nargs='+', help='parameters', action='append') args = parser.parse_args() if args.functions is None: print help(postprocess_operation) return params = ParamParser(args.param).run() params = [{}, {}] # FIXME holder.args = args caller(args.input, args.labels, args.output, args.functions, params) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!C&-{celltk/postprocess_operation.pyfrom utils.traces import TracesController from utils.pairwise import one_to_one_assignment, one_to_two_assignment import numpy as np from utils.traces import construct_traces_based_on_next, convert_traces_to_storage, label_traces np.random.seed(0) def gap_closing(cells, DISPLACEMENT=100, MASSTHRES=0.15, maxgap=4): ''' Connect cells between non-consecutive frames if it meets criteria. maxgap (int): the maximum frames allowed to connect two cells. ''' traces = construct_traces_based_on_next(cells) trhandler = TracesController(traces) # make sure not to have a cell as both disappered and appeared cells store_singleframe = [] for trace in trhandler.traces[:]: if len(trace) < 2: trhandler.traces.remove(trace) store_singleframe.append(trace) dist = trhandler.pairwise_dist() massdiff = trhandler.pairwise_mass() framediff = trhandler.pairwise_frame() withinarea = dist < DISPLACEMENT inmass = abs(massdiff) < MASSTHRES inframe = (framediff > 1) * (framediff <= maxgap) withinarea_inframe = withinarea * inframe * inmass # CHECK: distance as a fine cost withinarea_inframe = one_to_one_assignment(withinarea_inframe, dist) if withinarea_inframe.any(): disapp_idx, app_idx = np.where(withinarea_inframe) dis_cells = trhandler.disappeared() app_cells = trhandler.appeared() for disi, appi in zip(disapp_idx, app_idx): dis_cell, app_cell = dis_cells[disi], app_cells[appi] dis_cell.nxt = app_cell # You can simply reconstruct the trace, but here to reduce the calculation, # connect them explicitly. dis_trace = [i for i in trhandler.traces if dis_cell in i][0] app_trace = [i for i in trhandler.traces if app_cell in i][0] dis_trace.extend(trhandler.traces.pop(trhandler.traces.index(app_trace))) traces = traces + store_singleframe traces = label_traces(trhandler.traces) return convert_traces_to_storage(traces) def cut_short_traces(cells, minframe=4): ''' ''' if max([i.frame for i in cells]) < minframe: print "minframe set to the maximum" minframe = max([i.frame for i in cells]) '''handle division''' def list_parent_daughters(cells): cc = [(i.parent, i.label) for i in cells if i.parent is not None] parents = set([i[0] for i in cc]) parents = list(parents) store = [] for pt in parents: daughters = [i[1] for i in cc if i[0] == pt] store.append([pt] + daughters) return store pdsets = list_parent_daughters(cells) traces = construct_traces_based_on_next(cells) for pdset in pdsets: p0 = traces.pop([n for n, i in enumerate(traces) if pdset[0] == i[-1].label][0]) d0 = traces.pop([n for n, i in enumerate(traces) if pdset[1] == i[0].label][0]) d1 = traces.pop([n for n, i in enumerate(traces) if pdset[2] == i[0].label][0]) traces.append(p0 + d0) traces.append(p0 + d1) ''' Calculate the largest frame differences so it will go well with gap closing''' store = [] for trace in traces: frames = [i.frame for i in trace] if max(frames) - min(frames) >= minframe: store.append(trace) return convert_traces_to_storage(store) def detect_division(cells, DISPLACEMENT=50, maxgap=4, DIVISIONMASSERR=0.15): ''' ''' traces = construct_traces_based_on_next(cells) trhandler = TracesController(traces) store_singleframe = [] for trace in trhandler.traces[:]: if len(trace) < 2: trhandler.traces.remove(trace) store_singleframe.append(trace) dist = trhandler.pairwise_dist() massdiff = trhandler.pairwise_mass() framediff = trhandler.pairwise_frame() half_massdiff = massdiff + 0.5 withinarea = dist < DISPLACEMENT inframe = (framediff <= maxgap) * (framediff >= 1) halfmass = abs(half_massdiff) < DIVISIONMASSERR withinarea_inframe_halfmass = withinarea * inframe * halfmass # CHECK: now based on distance. par_dau = one_to_two_assignment(withinarea_inframe_halfmass, half_massdiff) # CHECK: If only one daughter is found ignore it. par_dau[par_dau.sum(axis=1) == 1] = False dis_cells = trhandler.disappeared() app_cells = trhandler.appeared() if par_dau.any(): disapp_idx, app_idx = np.where(par_dau) for disi, appi in zip(disapp_idx, app_idx): dis_cell = dis_cells[disi] app_cell = app_cells[appi] app_cell.parent = dis_cell.label # dis_cell.nxt = app_cell return convert_traces_to_storage(trhandler.traces + store_singleframe) PK!celltk/preprocess.py""" Any operations to make img from img. python celltk/preprocess.py -f gaussian_laplace -i c0/img_00000000* """ # from scipy.ndimage import imread import argparse from utils.file_io import make_dirs, imsave from utils.util import imread from utils.parser import ParamParser, parse_image_files import logging from utils.global_holder import holder import preprocess_operation logger = logging.getLogger(__name__) def caller(inputs, output, functions, params): holder.inputs = inputs make_dirs(output) logger.info("Functions {0} for {1} images.".format(functions, len(inputs))) for holder.frame, holder.path in enumerate(inputs): img = imread(holder.path) for function, param in zip(functions, params): func = getattr(preprocess_operation, function) img = func(img, **param) imsave(img, output, holder.path) logger.info("\tframe {0} done.".format(holder.frame)) def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--input", help="images", nargs="*") parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="*") parser.add_argument('-p', '--param', nargs='+', help='parameters', action='append') # parser.add_argument("-p", "--param", nargs="*", help="parameters", default=[]) args = parser.parse_args() if args.functions is None: print help(preprocess_operation) return params = ParamParser(args.param).run() args.input = parse_image_files(args.input) holder.args = args caller(args.input, args.output, args.functions, params) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!n{[$[$celltk/preprocess_operation.pyfrom __future__ import division from utils.filters import calc_lapgauss import SimpleITK as sitk import numpy as np from utils.preprocess_utils import homogenize_intensity_n4 from utils.preprocess_utils import convert_positive, estimate_background_prc from utils.preprocess_utils import resize_img from utils.preprocess_utils import histogram_matching, wavelet_subtraction_hazen, rolling_ball_subtraction_hazen from utils.filters import adaptive_thresh from utils.cp_functions import align_cross_correlation, align_mutual_information from utils.util import imread from glob import glob from utils.filters import interpolate_nan from scipy.optimize import minimize import logging from utils.global_holder import holder from utils.mi_align import calc_jitters_multiple, calc_crop_coordinates from utils.shading_correction import retrieve_ff_ref from scipy.ndimage.filters import gaussian_filter logger = logging.getLogger(__name__) np.random.seed(0) def gaussian_blur(img, SIGMA=3): img = gaussian_filter(img, sigma=SIGMA) return img def gaussian_laplace(img, SIGMA=2.5, NEG=False, OFFSET=0): if NEG: img = -calc_lapgauss(img, SIGMA) img[img < 0 ] = 0 img += OFFSET return img return calc_lapgauss(img, SIGMA) def curvature_anisotropic_smooth(img, NITER=10): fil = sitk.CurvatureAnisotropicDiffusionImageFilter() fil.SetNumberOfIterations(NITER) simg = sitk.GetImageFromArray(img.astype(np.float32)) sres = fil.Execute(simg) return sitk.GetArrayFromImage(sres) def background_subtraction_wavelet_hazen(img, THRES=100, ITER=5, WLEVEL=6, OFFSET=50): """Wavelet background subtraction. """ back = wavelet_subtraction_hazen(img.astype(np.float), ITER=ITER, THRES=THRES, WLEVEL=WLEVEL) img = img - back return convert_positive(img, OFFSET) def rolling_ball(img, RADIUS=100, SIGMA=3, OFFSET=50): """Rolling ball background subtraction. """ back = rolling_ball_subtraction_hazen(img.astype(np.float), RADIUS) img = img - back return convert_positive(img, OFFSET) def n4_illum_correction(img, RATIO=1.5, FILTERINGSIZE=50): """ Implementation of the N4 bias field correction algorithm. Takes some calculation time. It first calculates the background using adaptive_thesh. """ bw = adaptive_thresh(img, R=RATIO, FILTERINGSIZE=FILTERINGSIZE) img = homogenize_intensity_n4(img, ~bw) return img def n4_illum_correction_downsample(img, DOWN=2, RATIO=1.05, FILTERINGSIZE=50, OFFSET=10): """Faster but more insensitive to local illum bias. """ fil = sitk.ShrinkImageFilter() cc = sitk.GetArrayFromImage(fil.Execute(sitk.GetImageFromArray(img), [DOWN, DOWN])) bw = adaptive_thresh(cc, R=RATIO, FILTERINGSIZE=FILTERINGSIZE/DOWN) himg = homogenize_intensity_n4(cc, ~bw) himg = cc - himg # himg[himg < 0] = 0 bias = resize_img(himg, img.shape) img = img - bias return convert_positive(img, OFFSET) def align(img, CROP=0.05): """ CROP (float): crop images beforehand. When set to 0.05, 5% of each edges are cropped. """ if not hasattr(holder, "align"): if isinstance(holder.inputs[0], list) or isinstance(holder.inputs[0], tuple): inputs = [i[0] for i in holder.inputs] else: inputs = holder.inputs img0 = imread(inputs[0]) (ch, cw) = [int(CROP * i) for i in img0.shape] ch = None if ch == 0 else ch cw = None if cw == 0 else cw jitters = calc_jitters_multiple(inputs, ch, cw) holder.align = calc_crop_coordinates(jitters, img0.shape) logger.debug('holder.align set to {0}'.format(holder.align)) jt = holder.align[holder.frame] logger.debug('Jitter: {0}'.format(jt)) if img.ndim == 2: return img[jt[0]:jt[1], jt[2]:jt[3]] if img.ndim == 3: return img[jt[0]:jt[1], jt[2]:jt[3], :] def flatfield_references(img, ff_paths=['Pos0/img00.tif', 'Pos1/img01.tif'], exp_corr=False): """ Use empty images for background subtraction and illumination bias correction. Given multiple reference images, it will calculate median profile and use it for subtraction. If flatfield image has the same illumination pattern but different exposure to the img, turning on bg_align would calculate correction factor. ff_paths (str or List(str)): image path for flat fielding references. It can be single, multiple or path with wildcards. e.g. ff_paths = "FF/img_000000000_YFP*" ff_paths = ["FF/img_01.tif", "FF/img_02.tif"] """ store = [] if isinstance(ff_paths, str): ff_paths = [ff_paths, ] for i in ff_paths: for ii in glob(i): store.append(ii) ff_store = [] for path in store: ff_store.append(imread(path)) ff = np.median(np.dstack(ff_store), axis=2) if exp_corr: """If a reference is taken at different exposure, or exposure is not stable over time, this will try to correct for it. Majority of image needs to be a backrgound. """ def minimize_bg(img, ff, corr, perctile=50, weight=10): thres = np.percentile(img, perctile) res = img - corr * ff res = res[res < thres] return np.sum(res[res>0]) - weight * np.sum(res[res < 0]) """avoid having negative values yet suppress positive values in background region. """ func = lambda x: minimize_bg(img, ff, x) if not hasattr(holder, 'bg_corr'): holder.bg_corr = 1.0 ret = minimize(func, x0=holder.bg_corr, bounds=((0, None),)) holder.bg_corr = ret.x ff = ret.x * ff img = img - ff img[img < 0] = np.nan img = interpolate_nan(img) return img def histogram_match(img, BINS=1000, QUANT=100, THRES=False): """ If an optical system is not stable and shows global intensity changes over time, use this method to correct for it. Typically use for nuclear marker, where intensity and its patterns should be stable over time. """ if holder.frame == 0: holder.first_img = img else: img = histogram_matching(img, holder.first_img, BINS, QUANT, THRES) return img def shading_correction(img, ch='DAPI', whiteurl='http://archive.simtk.org/ktrprotocol/temp/ffref_20x3bin.npz', darkurl='http://archive.simtk.org/ktrprotocol/temp/ffdarkref_20x3bin.npz'): """only access if it has not""" if hasattr(holder, 'sc_ref'): if ch in holder.sc_ref: ref = holder.sc_ref[ch] darkref = holder.sc_dref[ch] else: ref, darkref = retrieve_ff_ref(whiteurl, darkurl) holder.sc_ref[ch] = ref holder.sc_dref[ch] = darkref else: ref, darkref = retrieve_ff_ref(whiteurl, darkurl) holder.sc_ref, holder.sc_dref = {}, {} holder.sc_ref[ch] = ref holder.sc_dref[ch] = darkref def correct_shade(img, ref, darkref, ch): img = img.astype(np.float) d0 = img.astype(np.float) - darkref[ch] d1 = ref[ch] - darkref[ch] d1[d1 < 0] = 0 return d1.mean() * d0/d1 img = correct_shade(img, ref, darkref, ch) return img def background_subtraction_wavelet(img, level=7, OFFSET=10): ''' It might be radical but works in many cases in terms of segmentation. Use "background_subtraction_wavelet_hazen" for a proper implementation. ''' from pywt import WaveletPacket2D from skimage.transform import resize def wavelet_subtraction(img, level): """6- 7 level is recommended""" if level == 0: return img wp = WaveletPacket2D(data=img.astype(np.uint16), wavelet='haar', mode='sym') back = resize(np.array(wp['a'*level].data), img.shape, order=3, mode='reflect')/(2**level) img = img - back return img img = wavelet_subtraction(img, level) return convert_positive(img, OFFSET) def np_arithmetic(img, npfunc='max'): func = getattr(np, npfunc) return func(img, axis=2) def stitch_images(img, POINTS=[(0,0),(0,0),(0,0),(0,0)]): from utils.stitch_utils import relative_position, stitching ''' Stitch images with 'Fiji/Stitch_image_Grid_Sequence' results. ''' rp = relative_position(POINTS) img = stitching(img, rp) img = np_arithmetic(img, 'max') return img def deep_unet(img, weight_path, region=1): """ Generates a probability map of cells using the UNet algorithm. Args: img (numpy.ndarray): image that will be segmented weight_path (string): path to weights file in .hdf5 format, can either bea local path or url. region (int): determines if each image will be saved individually (region =1) or setting region as None will save a stack of float32 images. Returns: pimg (numpy.ndarray): probability map image """ from utils.unet_predict import predict from utils.file_io import LocalPath from utils.global_holder import holder with LocalPath(weight_path) as wpath: pimg = predict(holder.path, wpath) pimg = np.moveaxis(pimg, 0, -1) return pimg[:, :, region] PK!ޤ celltk/segment.py""" python celltk/segment.py -i ~/covertrack/data/testimages/img_0000000* -f example_thres -o c1 -p THRES=2000 """ # from scipy.ndimage import imread import argparse import tifffile as tiff from os.path import basename, join import numpy as np import segment_operation from skimage.segmentation import clear_border from utils.filters import gray_fill_holes from skimage.morphology import remove_small_objects from utils.filters import label from scipy.ndimage.morphology import binary_opening from utils.util import imread from utils.file_io import make_dirs, imsave from utils.parser import ParamParser, parse_image_files from utils.global_holder import holder import logging logger = logging.getLogger(__name__) radius = [3, 100] def clean_labels(labels, rad, OPEN=3): """default cleaning. Fill holes, remove small and large objects and opening. """ labels = gray_fill_holes(labels) labels = clear_border(labels, buffer_size=2) labels = remove_small_objects(labels, rad[0]**2 * np.pi, connectivity=4) antimask = remove_small_objects(labels, rad[1]**2 * np.pi, connectivity=4) labels[antimask > 0] = False labels = label(binary_opening(labels, np.ones((int(OPEN), int(OPEN))), iterations=1)) return labels def caller(inputs, output, functions, params): make_dirs(output) logger.info("Functions {0} for {1} images.".format(functions, len(inputs))) for holder.frame, path in enumerate(inputs): holder.path = path img = imread(path) for function, param in zip(functions, params): func = getattr(segment_operation, function) img = func(img, **param) if isinstance(path, list) or isinstance(path, tuple): path = path[0] labels = clean_labels(img, radius) imsave(labels, output, path, dtype=np.int16) logger.info("\tframe {0}: {1} objects segmented.".format(holder.frame, len(np.unique(labels)))) def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--input", help="images", nargs="*") parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="*", default=None) parser.add_argument('-p', '--param', nargs='+', help='parameters', action='append') args = parser.parse_args() params = ParamParser(args.param).run() if args.functions is None: print help(segment_operation) return holder.args = args args.input = parse_image_files(args.input) caller(args.input, args.output, args.functions, params) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!wn  celltk/segment_operation.pyfrom __future__ import division from skimage.measure import label from utils.filters import adaptive_thresh from skimage.filters import threshold_otsu from utils.filters import label_watershed from utils.binary_ops import grey_dilation import numpy as np from scipy.ndimage import gaussian_laplace, binary_dilation, binary_opening, binary_closing np.random.seed(0) def constant_thres(img, THRES=2000, NEG=False): """take pixel above THRES as a foreground. Examples: >>> img = np.zeros((3, 3)) >>> img[0, 0] = 10 >>> img[2, 2] = 10 >>> example_thres(img, None, THRES=5) array([[1, 0, 0], [0, 0, 0], [0, 0, 2]]) """ if NEG: return label(img < THRES) return label(img > THRES) def global_otsu(img): global_thresh = threshold_otsu(img) return label(img > global_thresh) def adaptive_thres(img, FIL1=10, R1=100): """adaptive thresholding for picking objects with different brightness. """ bw = adaptive_thresh(img, R=R1, FILTERINGSIZE=FIL1) return label(bw) def adaptive_thres_two(img, FIL1=10, FIL2=100, R1=100, R2=100): """adaptive thresholding for picking objects with different brightness. FIL2 and R2 for removing background. """ bw = adaptive_thresh(img, R=R1, FILTERINGSIZE=FIL1) foreground = adaptive_thresh(img, R=R2, FILTERINGSIZE=FIL2) bw[-foreground] = 0 return label(bw) def adaptive_thres_otsu(img, FIL1=4, R1=1): """adaptive thresholding for picking objects with different brightness. Use Otsu's method for removing background """ bw = adaptive_thresh(img, R1, FIL1) foreground = global_otsu(img) > 0 bw[-foreground] = 0 return label(bw) def watershed_labels(labels, REG=10): """watershed to separate objects with concavity. Does not use intensity information but shape. """ return label_watershed(labels, regmax=REG) def lap_peak_local(img, separation=10, percentile=64, min_sigma=2, max_sigma=5, num_sigma=10): sigma_list = np.linspace(min_sigma, max_sigma, num_sigma) gl_images = [-gaussian_laplace(img, s) * s ** 2 for s in sigma_list] image_cube = np.dstack(gl_images) max_image = np.max(image_cube, axis=2) coords = grey_dilation(max_image, separation=separation, percentile=percentile) def mark_pos(im, coords): temp = np.zeros(im.shape) for c0, c1 in coords: temp[c0, c1] = 1 return temp bw = mark_pos(img, coords) return label(binary_dilation(bw, np.ones((3, 3)))) PK!-TOcelltk/subdetect.py""" python celltk/subdetect.py -l c3/img_00000000* -f ring_dilation -o c4 -p MARGIN=0 """ from utils.util import imread import tifffile as tiff import argparse from os.path import basename, join import numpy as np import subdetect_operation from itertools import izip_longest from utils.file_io import make_dirs, imsave, lbread from utils.parser import ParamParser from utils.global_holder import holder import logging logger = logging.getLogger(__name__) def caller(inputs, inputs_labels, output, functions, params): make_dirs(output) logger.info("Functions {0} for {1} images.".format(functions, len(inputs))) img = None for holder.frame, (path, pathl) in enumerate(izip_longest(inputs, inputs_labels)): if path is not None: img = imread(path) labels0 = lbread(pathl) for function, param in zip(functions, params): func = getattr(subdetect_operation, function) if img is not None: labels = func(labels0, img, **param) else: labels = func(labels0, **param) imsave(labels, output, pathl, dtype=np.int16) logger.info("\tframe {0} done.".format(holder.frame)) def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--input", help="images", nargs="*", default=[]) parser.add_argument("-l", "--labels", help="labels", nargs="+", default=[]) parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="+") parser.add_argument('-p', '--param', nargs='+', help='parameters', action='append') args = parser.parse_args() if args.functions is None: print help(subdetect_operation) return params = ParamParser(args.param).run() holder.args = args caller(args.input, args.labels, args.output, args.functions, params) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!?p p celltk/subdetect_operation.pyfrom utils.subdetect_utils import dilate_to_cytoring, dilate_to_cytoring_buffer from utils.concave_seg import levelset_geo_separete from utils.concave_seg import run_concave_cut from utils.filters import MultiSnakesCombined from utils.concave_seg import levelset_lap from utils.filters import label, adaptive_thresh from scipy.ndimage.filters import minimum_filter import numpy as np from scipy.ndimage import morphology from skimage.morphology import remove_small_objects from utils.labels_handling import convert_labels from utils.subdetect_utils import label_high_pass, label_nearest np.random.seed(0) def ring_dilation(labels, MARGIN=0, RINGWIDTH=4): """Create a ring around label. :param RINGWIDTH (int): Width of rings :param MARGIN (int): A region of rings is ignored if they are within MARGIN pixels away from label. Examples: >>> arr = np.zeros((5, 5));arr[2, 2] = 10 >>> ring_dilation(None, arr, None, MARGIN=1, RINGWIDTH=2) array([[ 0, 10, 10, 10, 0], [10, 0, 0, 0, 10], [10, 0, 0, 0, 10], [10, 0, 0, 0, 10], [ 0, 10, 10, 10, 0]], dtype=uint16) """ return dilate_to_cytoring(labels, RINGWIDTH, MARGIN) def ring_dilation_buffer(labels, MARGIN=0, RINGWIDTH=4, BUFFER=2): return dilate_to_cytoring_buffer(labels, RINGWIDTH, MARGIN, BUFFER) def ring_dilation_above_thres(labels, img, MARGIN=2, RINGWIDTH=4, EXTRA_RINGWIDTH=15, THRES=50): sub_label = dilate_to_cytoring(labels, RINGWIDTH, MARGIN) extra_sub_label = dilate_to_cytoring(labels, EXTRA_RINGWIDTH, RINGWIDTH) extra_sub_label[img < THRES] = 0 return sub_label + extra_sub_label def ring_dilation_above_offset_buffer(labels, img, MARGIN=0, RINGWIDTH=2, BUFFER=2, OFFSET=100, FILSIZE=50): """Dilate from label to make a ring. Calculate the local minimum as a background, and if image is less brighter than background + offset, remove the region from the ring. """ sub_label = dilate_to_cytoring_buffer(labels, RINGWIDTH, MARGIN, BUFFER) minimg = minimum_filter(img, size=FILSIZE) sub_label[img < (minimg + OFFSET)] = 0 return sub_label def ring_dilation_above_adaptive(labels, img, MARGIN=0, RINGWIDTH=4, BUFFER=2, RATIO=1.05, FILSIZE=10): sub_labels = dilate_to_cytoring_buffer(labels, RINGWIDTH, MARGIN, BUFFER) bw = adaptive_thresh(img, R=RATIO, FILTERINGSIZE=FILSIZE) sub_labels[-bw] = 0 return sub_labels def geodesic_levelset(labels, img, NITER=10, PROP=1): """propagated outwards until it sticks to the shape boundaries in an image. Generally used for making/repairing too small objects bigger. Larger NITER will give more propagation and roundy object. """ return levelset_geo_separete(img, labels, niter=NITER, prop=PROP) def concave_cut(labels, img, SMALL_RAD=7, LARGE_RAD=14, EDGELEN=6, THRES=180): """ Attempt a cut for objects larger than np.pi * large_rad ** 2. For each pixel, the angle of two vectors pointing to boundary pixels distant by EDGELEN is calculated to find strong concavity more than THRES angle. The cut line is chosen by minimizing intensity below a line between two pixels. To reduce calculation costs, only pixels on a watershed line are assessed. SMALL_RAD (int): minimum radius of nuclei LARGE_RAD (int): maximum radius of nuclei EDGELEN (int): length of triangle edges to calculate angle THRES (int): threshold for concavity angles """ labels = run_concave_cut(img, labels, small_rad=SMALL_RAD, large_rad=LARGE_RAD, EDGELEN=EDGELEN, THRES=THRES) return labels def watershed_cut(labels, img, MIN_SIGMA=2, MAX_SIGMA=10, THRES=1000): from utils.filters import lap_local_max, sitk_watershed_intensity sigma_list = range(int(MIN_SIGMA), int(MAX_SIGMA)) local_maxima = lap_local_max(img, sigma_list, THRES) return sitk_watershed_intensity(labels, local_maxima) def propagate_multisnakes(labels, img, NITER=3, SMOOTHING=1, lambda1=1, lambda2=1): """ Higher lambda2 relative to lambda1 gives more outward propagation. """ ms = MultiSnakesCombined(img, labels, smoothing=SMOOTHING, lambda1=lambda1, lambda2=lambda2) labels = ms.multi_step(niter=NITER) return labels def laplacian_levelset(labels, img, NITER=100, CURVE=3, PROP=-1): return label(levelset_lap(img, labels, NITER, CURVE, PROP)) def voronoi_cut(labels): from utils.subdetect_utils import voronoi_expand return voronoi_expand(labels) def detect_puncta_voronoi(labels, img, level=7, PERC=50, FILSIZE=1): from utils.subdetect_utils import voronoi_expand from utils.fish_detect import detect_puncta vor = voronoi_expand(labels) puncta = detect_puncta(img, level=level, PERC=PERC, FILSIZE=FILSIZE) vor[puncta == 0] = 0 return vor def morphological(labels, func='grey_opening', size=3, iterations=1): morph_operation = getattr(morphology, func) for i in range(iterations): labels = morph_operation(labels, size=(size, size)) return labels def watershed_divide(labels, regmax=10, min_size=100): """ divide objects in labels with watershed segmentation. regmax: min_size: objects smaller than this size will not be divided. """ from utils.subdetect_utils import watershed_labels large_labels = remove_small_objects(labels, min_size, connectivity=4) labels[large_labels > 0] = 0 ws_large = watershed_labels(large_labels, regmax) ws_large += labels.max() ws_large[ws_large == labels.max()] = 0 return labels + ws_large def cytoplasm_levelset(labels, img, niter=20, dt=-0.5, thres=0.5): """ Segment cytoplasm from supplied nuclear labels and probability map Expand using level sets method from nuclei to membrane. Uses an implementation of Level set method. See: https://wiseodd.github.io/techblog/2016/11/05/levelset-method/ Args: labels (numpy.ndarray): nuclear mask labels img (numpy.ndarray): probability map niter (int): step size to expand mask, number of iterations to run the levelset algorithm dt (float): negative values for porgation, positive values for shrinking thres (float): threshold of probability value to extend the nuclear mask to Returns: cytolabels (numpy.ndarray): cytoplasm mask labels """ from skimage.morphology import closing, disk, remove_small_holes from utils.dlevel_set import dlevel_set phi = labels.copy() phi[labels == 0] = 1 phi[labels > 0] = -1 outlines = img.copy() outlines = -outlines outlines = outlines - outlines.min() outlines = outlines/outlines.max() mask = outlines < thres phi = dlevel_set(phi, outlines, niter=niter, dt=dt, mask=mask) cytolabels = label(remove_small_holes(label(phi < 0))) cytolabels = closing(cytolabels, disk(3)) temp = cytolabels.copy() temp[labels == 0] = 0 cytolabels = convert_labels(temp, labels, cytolabels) return cytolabels def segment_bacteria(nuc, img, slen=3, SIGMA=0.5,THRES=100, CLOSE=20, THRESCHANGE=1000, MINAREA=5): """ Segment bacteria and assign to closest nucleus Args: nuc (numpy.ndarray): nuclear mask labels img (numpy.ndarray): image in bacterial channel slen (int): Size of Gaussian kernel SIGMA (float): Standard deviation for Gaussian kernel THRES (int): Threshold pixel intensity fo real signal CLOSE (int): Radius for disk used to return morphological closing of the image (dilation followed by erosion to remove dark spots and connect bright cracks) THRESCHANGE (int): argument unnecessary? MINAREA (int): minimum area in pixels for a bacterium Returns: labels (numpy.ndarray[np.uint16]): bacterial mask labels """ labels = label_high_pass(img, slen=slen, SIGMA=SIGMA, THRES=THRES, CLOSE=3) if labels.any(): labels, comb, nuc_prop, nuc_loc = label_nearest(img, labels, nuc) from skimage.morphology import remove_small_objects labels = remove_small_objects(labels, MINAREA) return labels.astype(np.uint16) PK!Rަp p celltk/track.py""" python celltk/tracking.py -f nearest_neighbor -i c0/img_00000000* -l c1/img_0000000 0* -p DISPLACEMENT=10 MASSTHRES=0.2 """ from utils.util import imread import argparse import tifffile as tiff from os.path import basename, join import numpy as np import os import track_operation from utils.file_io import make_dirs, imsave, lbread from utils.parser import ParamParser from utils.global_holder import holder import logging logger = logging.getLogger(__name__) def neg2poslabels(labels): if not hasattr(holder, 'max'): holder.max = labels.max() holder.max = max(holder.max, labels.max()) negatives = np.unique(labels[labels < 0]) for i in negatives: holder.max += 1 labels[labels == i] = holder.max return labels def caller(inputs, inputs_labels, output, functions, params): make_dirs(output) img0, labels0 = imread(inputs[0]), lbread(inputs_labels[0]).astype(np.int16) labels0 = neg2poslabels(labels0) imsave(labels0, output, basename(inputs[0]), dtype=np.int16) for holder.frame, (path, pathl) in enumerate(zip(inputs[1:], inputs_labels[1:])): img1, labels1 = imread(path), lbread(pathl) labels1 = -labels1 for fnum, (function, param) in enumerate(zip(functions, params)): func = getattr(track_operation, function) if not (labels1 < 0).any(): continue labels0, labels1 = func(img0, img1, labels0, -labels1, **param) logger.debug('\t{0} with {1}: {2}'.format(function, param, len(set(labels1[labels1 < 0])))) logger.info("\tframe {0}: {1} objects linked and {2} unlinked.".format(holder.frame, len(set(labels1[labels1 > 0])), len(set(labels1[labels1 < 0])))) labels0 = neg2poslabels(labels1) img0 = img1 imsave(labels0, output, path, dtype=np.int16) def main(): parser = argparse.ArgumentParser() parser.add_argument("-i", "--input", help="images", nargs="+") parser.add_argument("-l", "--labels", help="labels", nargs="+") parser.add_argument("-o", "--output", help="output directory", type=str, default='temp') parser.add_argument("-f", "--functions", help="functions", nargs="+") parser.add_argument('-p', '--param', nargs='+', help='parameters', action='append') args = parser.parse_args() if args.functions is None: print help(track_operation) return params = ParamParser(args.param).run() holder.args = args caller(args.input, args.labels, args.output, args.functions, params) if __name__ == "__main__": logging.basicConfig(level=logging.INFO) main() PK!27))celltk/track_operation.py""" Label labels1 to the same value from labels0. Not tracked is negative. Turn labels0 into negative. """ from utils.filters import label from utils.postprocess_utils import regionprops from scipy.spatial.distance import cdist from utils.track_utils import calc_massdiff, find_one_to_one_assign from utils.global_holder import holder # holder is used to store parameters that needs to be called many times import numpy as np from utils.track_utils import call_lap, pick_closer_cost from utils.filters import labels2outlines from utils.concave_seg import wshed_raw, CellCutter from utils.track_utils import _find_best_neck_cut, _update_labels_neck_cut from utils.global_holder import holder from scipy.ndimage import gaussian_laplace, binary_dilation from utils.binary_ops import grey_dilation import logging logger = logging.getLogger(__name__) np.random.seed(0) def nearest_neighbor(img0, img1, labels0, labels1, DISPLACEMENT=20, MASSTHRES=0.2): """ labels0 and labels1: the positive values for non-tracked objects and the negative values for tracked objects. """ labels = -labels1.copy() rps0 = regionprops(labels0, img0) rps1 = regionprops(labels1, img1) if not rps0 or not rps1: return labels0, labels dist = cdist([i.centroid for i in rps0], [i.centroid for i in rps1]) massdiff = calc_massdiff(rps0, rps1) binary_cost = (dist < DISPLACEMENT) * (abs(massdiff) < MASSTHRES) idx1, idx0 = find_one_to_one_assign(binary_cost) for i0, i1 in zip(idx0, idx1): labels[labels1 == rps1[i1].label] = rps0[i0].label labels0[labels0 == rps0[i0].label] = -rps0[i0].label return labels0, labels def nn_closer(img0, img1, labels0, labels1, DISPLACEMENT=30, MASSTHRES=0.25): labels = -labels1.copy() rps0 = regionprops(labels0, img0) rps1 = regionprops(labels1, img1) if not rps0 or not rps1: return labels0, labels dist = cdist([i.centroid for i in rps0], [i.centroid for i in rps1]) massdiff = calc_massdiff(rps0, rps1) binary_cost = (dist < DISPLACEMENT) * (abs(massdiff) < MASSTHRES) binary_cost = pick_closer_cost(binary_cost, dist) binary_cost = pick_closer_cost(binary_cost.T, dist.T).T idx1, idx0 = find_one_to_one_assign(binary_cost) for i0, i1 in zip(idx0, idx1): labels[labels1 == rps1[i1].label] = rps0[i0].label labels0[labels0 == rps0[i0].label] = -rps0[i0].label return labels0, labels def run_lap(img0, img1, labels0, labels1, DISPLACEMENT=30, MASSTHRES=0.2): '''Linear assignment problem for mammalian cells. Cost matrix is simply the distance. costDie and costBorn are variables changing over frame. Update it through holder. If you have more than 1000 cells per image, it may cause a slowdown. Args: DISPLACEMENT (int): The maximum distance (in pixel) MASSTHRES (float): The maximum difference of total intensity changes. 0.2 means it allows for 20% total intensity changes. ''' labels = -labels1.copy() rps0 = regionprops(labels0, img0) rps1 = regionprops(labels1, img1) if not rps0 or not rps1: return labels0, labels dist = cdist([i.centroid for i in rps0], [i.centroid for i in rps1]) massdiff = calc_massdiff(rps0, rps1) '''search radius is now simply set by maximum displacement possible. In the future, I might add data-driven approach mentioned in LAP paper (supple pg.7)''' dist[dist > DISPLACEMENT] = np.Inf # assign a large cost for unlikely a pair # dist[abs(massdiff) > MASSTHRES] = np.Inf cost = dist if cost.shape[0] == 0 or cost.shape[1] == 0: return labels0, labels # Define initial costBorn and costDie in the first frame if not hasattr(holder, 'cost_born') or not hasattr(holder, 'cost_die'): holder.cost_born = np.percentile(cost[~np.isinf(cost)], 80) holder.cost_die = np.percentile(cost[~np.isinf(cost)], 80) # try: binary_cost = call_lap(cost, holder.cost_die, holder.cost_born) # The first assignment of np.Inf is to reduce calculation of linear assignment. # This part will make sure that cells outside of these range do not get connected. binary_cost[(np.abs(massdiff) > MASSTHRES)] = False binary_cost[(dist > DISPLACEMENT)] = False gp, gc = np.where(binary_cost) idx0, idx1 = list(gp), list(gc) for i0, i1 in zip(idx0, idx1): labels[labels1 == rps1[i1].label] = rps0[i0].label labels0[labels0 == rps0[i0].label] = -rps0[i0].label # update cost linked_dist = [dist[i0, i1] for i0, i1 in zip(idx0, idx1)] if linked_dist: cost = np.max(linked_dist)*1.05 if cost != 0: # solver freezes if cost is 0 holder.cost_born, holder.cost_die = cost, cost return labels0, labels def track_neck_cut(img0, img1, labels0, labels1, DISPLACEMENT=10, MASSTHRES=0.2, EDGELEN=5, THRES_ANGLE=180, WSLIMIT=False, SMALL_RAD=None, CANDS_LIMIT=300): """ Adaptive segmentation by using tracking informaiton. Separate two objects by making a cut at the deflection. For each points on the outline, it will make a triangle separated by EDGELEN and calculates the angle facing inside of concave. The majority of cells need to be tracked before the this method to calculate LARGE_RAD and SMALL_RAD. EDGELEN (int): A length of edges of triangle on the nuclear perimeter. THRES_ANGLE (int): Define the neck points if a triangle has more than this angle. STEPLIM (int): points of neck needs to be separated by at least STEPLIM in parimeters. WSLIMIT (bool): Limit search points to ones overlapped with watershed transformed images. Set it True if calculation is slow. SMALL_RAD (int or None): The smallest radius of candidate objects. If you have many cells, set it to None will infer the radius from previous frame. CANDS_LIMIT(int): use lower if slow. limit a number of searches. """ labels0, labels = nn_closer(img0, img1, labels0, labels1, DISPLACEMENT, MASSTHRES) labels1 = -labels.copy() if SMALL_RAD is None and not hasattr(holder, 'SMALL_RAD'): tracked_area = [i.area for i in regionprops(labels)] holder.SMALL_RAD = np.sqrt(np.percentile(tracked_area, 5)/np.pi) elif SMALL_RAD is not None: holder.SMALL_RAD = SMALL_RAD SMALL_RAD = holder.SMALL_RAD rps0 = regionprops(labels0, img0) unique_labels = np.unique(labels1) unique_labels = unique_labels[unique_labels > 0] if WSLIMIT: wlines = wshed_raw(labels1 > 0, img1) else: wlines = np.ones(labels1.shape, np.bool) store = [] coords_store = [] for label_id in unique_labels: if label_id == 0: continue cc = CellCutter(labels1 == label_id, img1, wlines, small_rad=SMALL_RAD, EDGELEN=EDGELEN, THRES=THRES_ANGLE, CANDS_LIMIT=CANDS_LIMIT) cc.prepare_coords_set() candidates = cc.search_cut_candidates(cc.bw.copy(), cc.coords_set[:CANDS_LIMIT]) for c in candidates: c.raw_label = label_id store.append(candidates) coords_store.append(cc.coords_set) coords_store = [i for i in coords_store if i] # Attempt a first cut. good_cells = _find_best_neck_cut(rps0, store, DISPLACEMENT, MASSTHRES) labels0, labels = _update_labels_neck_cut(labels0, labels1, good_cells) labels0, labels = nn_closer(img0, img1, labels0, -labels, DISPLACEMENT, MASSTHRES) while good_cells: rps0 = regionprops(labels0, img0) labels1 = -labels.copy() rps0 = regionprops(labels0, img0) unique_labels = np.unique(labels1) store = [] for label_id in unique_labels: if label_id == 0: continue bw = labels1 == label_id coords_set = [i for i in coords_store if bw[i[0][0][0], i[0][0][1]]] if not coords_set: continue coords_set = coords_set[0] candidates = cc.search_cut_candidates(bw, coords_set) for c in candidates: c.raw_label = label_id store.append(candidates) coords_store.append(coords_set) good_cells = _find_best_neck_cut(rps0, store, DISPLACEMENT, MASSTHRES) labels0, labels = _update_labels_neck_cut(labels0, labels1, good_cells) labels0, labels = nn_closer(img0, img1, labels0, -labels, DISPLACEMENT, MASSTHRES) return labels0, labels def watershed_distance(img0, img1, labels0, labels1, DISPLACEMENT=10, MASSTHRES=0.2, REGMAX=10, MIN_SIZE=50): ''' Adaptive segmentation by using tracking informaiton. watershed existing label, meaning make a cut at the deflection. After the cuts, objects will be linked if they are within DISPLACEMENT and MASSTHRES. If two candidates are found, it will pick a closer one. track_neck_cut may be more sensitive but it takes a long time if objects are not smooth. Args: REGMAX (int): Watershed seeds will be separated by at least REGMAX. Smaller REGMAX will allow more cuts. DISPLACEMENT (int): The maximum distance (in pixel) MASSTHRES (float): The maximum difference of total intensity changes. 0.2 means it allows for 20% total intensity changes. ''' labels0, labels = nn_closer(img0, img1, labels0, labels1, DISPLACEMENT, MASSTHRES) def _wd(labels0, labels, img0, img1): labels1 = -labels.copy() rps0 = regionprops(labels0, img0) from subdetect_operation import watershed_divide # DO NOT MOVE IT from utils.track_utils import _find_match untracked_labels = labels1.copy() untracked_labels[untracked_labels < 0] = 0 wshed_labels = watershed_divide(untracked_labels, regmax=REGMAX, min_size=MIN_SIZE) wshed_labels = label(wshed_labels) store = regionprops(wshed_labels, img1) good_cells = _find_match(rps0, store, DISPLACEMENT, MASSTHRES) for gc in good_cells: # needed to reuse _update_labels_neck_cut gccrds = gc.coords[0] gc.raw_label = labels1[gccrds[0], gccrds[1]] labels0, labels = _update_labels_neck_cut(labels0, labels1, good_cells) labels0, labels = nn_closer(img0, img1, labels0, -labels, DISPLACEMENT, MASSTHRES) return labels0, labels, good_cells labels0, labels, good_cells = _wd(labels0, labels, img0, img1) while good_cells: labels0, labels, good_cells = _wd(labels0, labels, img0, img1) return labels0, labels PK!celltk/utils/__init__.pyPK!PF celltk/utils/_model_builder.pyimport keras.layers import keras.models import tensorflow as tf CONST_DO_RATE = 0.5 option_dict_conv = {"activation": "relu", "border_mode": "same"} option_dict_bn = {"mode": 0, "momentum" : 0.9} # returns a core model from input to 64 channels of the same size def get_core(dim1, dim2, dim3): # assume dim1 x dim2 image with dim3 color channels x = keras.layers.Input(shape=(dim1, dim2, dim3)) a = keras.layers.Convolution2D(64, 3, 3, **option_dict_conv)(x) a = keras.layers.BatchNormalization(**option_dict_bn)(a) a = keras.layers.Convolution2D(64, 3, 3, **option_dict_conv)(a) a = keras.layers.BatchNormalization(**option_dict_bn)(a) y = keras.layers.MaxPooling2D()(a) b = keras.layers.Convolution2D(128, 3, 3, **option_dict_conv)(y) b = keras.layers.BatchNormalization(**option_dict_bn)(b) b = keras.layers.Convolution2D(128, 3, 3, **option_dict_conv)(b) b = keras.layers.BatchNormalization(**option_dict_bn)(b) y = keras.layers.MaxPooling2D()(b) c = keras.layers.Convolution2D(256, 3, 3, **option_dict_conv)(y) c = keras.layers.BatchNormalization(**option_dict_bn)(c) c = keras.layers.Convolution2D(256, 3, 3, **option_dict_conv)(c) c = keras.layers.BatchNormalization(**option_dict_bn)(c) y = keras.layers.MaxPooling2D()(c) d = keras.layers.Convolution2D(512, 3, 3, **option_dict_conv)(y) d = keras.layers.BatchNormalization(**option_dict_bn)(d) d = keras.layers.Convolution2D(512, 3, 3, **option_dict_conv)(d) d = keras.layers.BatchNormalization(**option_dict_bn)(d) # UP d = keras.layers.UpSampling2D()(d) y = keras.layers.merge([d, c], concat_axis=3, mode="concat") e = keras.layers.Convolution2D(256, 3, 3, **option_dict_conv)(y) e = keras.layers.BatchNormalization(**option_dict_bn)(e) e = keras.layers.Convolution2D(256, 3, 3, **option_dict_conv)(e) e = keras.layers.BatchNormalization(**option_dict_bn)(e) e = keras.layers.UpSampling2D()(e) y = keras.layers.merge([e, b], concat_axis=3, mode="concat") f = keras.layers.Convolution2D(128, 3, 3, **option_dict_conv)(y) f = keras.layers.BatchNormalization(**option_dict_bn)(f) f = keras.layers.Convolution2D(128, 3, 3, **option_dict_conv)(f) f = keras.layers.BatchNormalization(**option_dict_bn)(f) f = keras.layers.UpSampling2D()(f) y = keras.layers.merge([f, a], concat_axis=3, mode="concat") y = keras.layers.Convolution2D(64, 3, 3, **option_dict_conv)(y) y = keras.layers.BatchNormalization(**option_dict_bn)(y) y = keras.layers.Convolution2D(64, 3, 3, **option_dict_conv)(y) y = keras.layers.BatchNormalization(**option_dict_bn)(y) return [x, y] def get_model(dim1, dim2, dim3, activation="softmax"): [x, y] = get_core(dim1, dim2, dim3) y = keras.layers.Convolution2D(3, 1, 1, **option_dict_conv)(y) if activation is not None: y = keras.layers.Activation(activation)(y) model = keras.models.Model(x, y) return modelPK!_celltk/utils/_munkres.pyimport numpy as np from scipy.optimize import linear_sum_assignment def munkres(arr): temp = np.zeros(arr.shape, np.bool) ind = linear_sum_assignment(arr) temp[ind] = True return tempPK!Usscelltk/utils/_mutinfo.py""" Replace np histogram to fast-histogram. """ # Copyright (C) 2013 Oskar Maier # # This program is free software: you can redistribute it and/or modify # it under the terms of the GNU General Public License as published by # the Free Software Foundation, either version 3 of the License, or # (at your option) any later version. # # This program is distributed in the hope that it will be useful, # but WITHOUT ANY WARRANTY; without even the implied warranty of # MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the # GNU General Public License for more details. # # You should have received a copy of the GNU General Public License # along with this program. If not, see . # # author Oskar Maier # version r0.1.0 # since 2013-07-09 # status Release # build-in modules # third-party modules import numpy from fast_histogram import histogram1d, histogram2d # code def mutual_information(i1, i2, bins=256): r""" Computes the mutual information (MI) (a measure of entropy) between two images. MI is not real metric, but a symmetric and nonnegative similarity measures that takes high values for similar images. Negative values are also possible. Intuitively, mutual information measures the information that ``i1`` and ``i2`` share: it measures how much knowing one of these variables reduces uncertainty about the other. The Entropy is defined as: .. math:: H(X) = - \sum_i p(g_i) * ln(p(g_i) with :math:`p(g_i)` being the intensity probability of the images grey value :math:`g_i`. Assuming two images :math:`R` and :math:`T`, the mutual information is then computed by comparing the images entropy values (i.e. a measure how well-structured the common histogram is). The distance metric is then calculated as follows: .. math:: MI(R,T) = H(R) + H(T) - H(R,T) = H(R) - H(R|T) = H(T) - H(T|R) A maximization of the mutual information is equal to a minimization of the joint entropy. Parameters ---------- i1 : array_like The first image. i2 : array_like The second image. bins : integer The number of histogram bins (squared for the joined histogram). Returns ------- mutual_information : float The mutual information distance value between the supplied images. Raises ------ ArgumentError If the supplied arrays are of different shape. """ # pre-process function arguments i1 = numpy.asarray(i1) i2 = numpy.asarray(i2) # # validate function arguments # if not i1.shape == i2.shape: # raise ArgumentError('the two supplied array-like sequences i1 and i2 must be of the same shape') # compute i1 and i2 histogram range i1_range = __range(i1, bins) i2_range = __range(i2, bins) # compute joined and separated normed histograms i1i2_hist = histogram2d(i1.flatten(), i2.flatten(), bins=bins, range=[i1_range, i2_range]) # Note: histogram2d does not flatten array on its own i1_hist = histogram1d(i1, bins=bins, range=i1_range) i2_hist = histogram1d(i2, bins=bins, range=i2_range) # compute joined and separated entropy i1i2_entropy = __entropy(i1i2_hist) i1_entropy = __entropy(i1_hist) i2_entropy = __entropy(i2_hist) # compute and return the mutual information distance return i1_entropy + i2_entropy - i1i2_entropy def __range(a, bins): '''Compute the histogram range of the values in the array a according to scipy.stats.histogram.''' a = numpy.asarray(a) a_max = a.max() a_min = a.min() s = 0.5 * (a_max - a_min) / float(bins - 1) return (a_min - s, a_max + s) def __entropy(data): '''Compute entropy of the flattened data set (e.g. a density distribution).''' # normalize and convert to float data = data/float(numpy.sum(data)) # for each grey-value g with a probability p(g) = 0, the entropy is defined as 0, therefore we remove these values and also flatten the histogram data = data[numpy.nonzero(data)] # compute entropy return -1. * numpy.sum(data * numpy.log2(data)) PK!FPcelltk/utils/binary_ops.pyimport numpy as np from scipy import ndimage from scipy.spatial import cKDTree import warnings def validate_tuple(value, ndim): if not hasattr(value, '__iter__'): return (value,) * ndim if len(value) == ndim: return tuple(value) raise ValueError("List length should have same length as image dimensions.") def drop_close(pos, separation, intensity=None): """ Adapted from trackpy """ """ Removes features that are closer than separation from other features. When intensity is given, the one with the lowest intensity is dropped: else the most topleft is dropped (to avoid randomness)""" to_drop = where_close(pos, separation, intensity) return np.delete(pos, to_drop, axis=0) def where_close(pos, separation, intensity=None): """ Adapted from trackpy """ """ Returns indices of features that are closer than separation from other features. When intensity is given, the one with the lowest intensity is returned: else the most topleft is returned (to avoid randomness)""" if len(pos) == 0: return [] separation = validate_tuple(separation, pos.shape[1]) if any([s == 0 for s in separation]): return [] # Rescale positions, so that pairs are identified below a distance # of 1. pos_rescaled = pos / separation duplicates = cKDTree(pos_rescaled, 30).query_pairs(1 - 1e-7) if len(duplicates) == 0: return [] index_0 = np.fromiter((x[0] for x in duplicates), dtype=int) index_1 = np.fromiter((x[1] for x in duplicates), dtype=int) if intensity is None: to_drop = np.where(np.sum(pos_rescaled[index_0], 1) > np.sum(pos_rescaled[index_1], 1), index_1, index_0) else: intensity = np.asarray(intensity) intensity_0 = intensity[index_0] intensity_1 = intensity[index_1] to_drop = np.where(intensity_0 > intensity_1, index_1, index_0) edge_cases = intensity_0 == intensity_1 if np.any(edge_cases): index_0 = index_0[edge_cases] index_1 = index_1[edge_cases] to_drop[edge_cases] = np.where(np.sum(pos_rescaled[index_0], 1) > np.sum(pos_rescaled[index_1], 1), index_1, index_0) return np.unique(to_drop) def grey_dilation(image, separation, percentile=64, margin=None, precise=True): """ Adapted and modified from trackpy. It takes images not integer types. """ """Find local maxima whose brightness is above a given percentile. Parameters ---------- image : ndarray For best performance, provide an integer-type array. If the type is not of integer-type, the image will be normalized and coerced to uint8. separation : number or tuple of numbers Minimum separation between maxima. See precise for more information. percentile : float in range of [0,100], optional Features must have a peak brighter than pixels in this percentile. This helps eliminate spurious peaks. Default 64. margin : integer or tuple of integers, optional Zone of exclusion at edges of image. Default is ``separation / 2``. precise : boolean, optional Determines whether there will be an extra filtering step (``drop_close``) discarding features that are too close. Degrades performance. Because of the square kernel used, too many features are returned when precise=False. Default True. See Also -------- drop_close : removes features that are too close to brighter features grey_dilation_legacy : local maxima finding routine used until trackpy v0.3 """ ndim = image.ndim separation = validate_tuple(separation, ndim) if margin is None: margin = tuple([int(s / 2) for s in separation]) # Compute a threshold based on percentile. threshold = np.percentile(image, percentile) if np.isnan(threshold): warnings.warn("Image is completely black.", UserWarning) return np.empty((0, ndim)) # Find the largest box that fits inside the ellipse given by separation size = [int(2 * s / np.sqrt(ndim)) for s in separation] # The intersection of the image with its dilation gives local maxima. dilation = ndimage.grey_dilation(image, size, mode='constant') maxima = (image == dilation) & (image > threshold) if np.sum(maxima) == 0: warnings.warn("Image contains no local maxima.", UserWarning) return np.empty((0, ndim)) pos = np.vstack(np.where(maxima)).T # Do not accept peaks near the edges. shape = np.array(image.shape) near_edge = np.any((pos < margin) | (pos > (shape - margin - 1)), 1) pos = pos[~near_edge] if len(pos) == 0: warnings.warn("All local maxima were in the margins.", UserWarning) return np.empty((0, ndim)) # Remove local maxima that are too close to each other if precise: pos = drop_close(pos, separation, image[maxima][~near_edge]) return pos PK!x;;celltk/utils/concave_seg.pyimport numpy as np from scipy.spatial.distance import cdist import SimpleITK as sitk from postprocess_utils import regionprops from filters import label import cv2 from skimage.draw import line from scipy.ndimage import binary_erosion from mahotas.segmentation import gvoronoi from skimage.segmentation import find_boundaries from skimage.morphology import thin from functools import partial from track_utils import calc_massdiff, find_one_to_one_assign from scipy.spatial.distance import cdist def cut_neck(template, r0, c0, r1, c1): """Given a label image and two coordinates, it will draw a line which intensity is 0. """ # rr, cc, _ = line_aa(r0, c0, r1, c1) rr, cc = line(r0, c0, r1, c1) template[rr, cc] = 0 return template def make_candidates(cl_label, s0, s1, e0, e1): cut_label = cut_neck(cl_label, s0, s1, e0, e1) cand_label = label(cut_label, connectivity=1) cand_rps = regionprops(cand_label) return cand_rps def make_candidates_img(cl_label, s0, s1, e0, e1, img): cut_label = cut_neck(cl_label, s0, s1, e0, e1) cand_label = label(cut_label, connectivity=1) cand_rps = regionprops(cand_label, img) return cand_rps def intensity_below_line(img, r0, c0, r1, c1): rr, cc = line(r0, c0, r1, c1) return img[rr, cc] def calc_shortest_step_coords(coords, co1, co2): co1 = [i for i, c in enumerate(coords) if (c == co1).all()][0] co2 = [i for i, c in enumerate(coords) if (c == co2).all()][0] return min(abs(co2 - co1), abs(co1 + coords.shape[0] - co2)) def find_oriented_coords(outline): cnt = cv2.findContours(outline.astype(np.uint8), cv2.RETR_TREE, cv2.CHAIN_APPROX_NONE)[1][0] cnt = np.flipud(cnt) # assert cv2.contourArea(cnt, oriented=True) > 0 if not cv2.contourArea(cnt, oriented=True) > 0: return None return np.fliplr(np.squeeze(cnt)) def keep_labels(ref_labels, conv_labels): from collections import Counter pairs = [] for i in [ii for ii in np.unique(conv_labels) if not ii == 0]: c = Counter(ref_labels[conv_labels == i]) pairs.append((i, c.most_common()[0][0])) labels = conv_labels.copy() for p1, p2 in pairs: labels[conv_labels == p1] = p2 return labels def levelset_lap(img, labels, NITER=100, CURVE=3, PROP=-1): simg = sitk.GetImageFromArray(img, sitk.sitkFloat32) seg = sitk.GetImageFromArray(labels) init_ls = sitk.SignedMaurerDistanceMap(seg, insideIsPositive=True, useImageSpacing=True) lsFilter = sitk.LaplacianSegmentationLevelSetImageFilter() lsFilter.SetMaximumRMSError(0.02) lsFilter.SetNumberOfIterations(NITER) lsFilter.SetCurvatureScaling(CURVE) lsFilter.SetPropagationScaling(PROP) lsFilter.ReverseExpansionDirectionOn() ls = lsFilter.Execute(init_ls, sitk.Cast(simg, sitk.sitkFloat32)) ls = label(sitk.GetArrayFromImage(ls) > 0) return keep_labels(labels, ls) def levelset_geo(img, labels, advec=3, propagation=0.75, niter=100): """Propagate outwards """ simg = sitk.GetImageFromArray(img, sitk.sitkFloat32) labels = label(binary_erosion(labels, np.ones((3, 3)))) seg = sitk.GetImageFromArray(labels) init_ls = sitk.SignedMaurerDistanceMap(seg, insideIsPositive=True, useImageSpacing=True) gfil = sitk.GeodesicActiveContourLevelSetImageFilter() gfil.SetMaximumRMSError(0.02) gfil.SetNumberOfIterations(niter) gfil.SetAdvectionScaling(advec) gfil.SetCurvatureScaling(1) gfil.SetPropagationScaling(propagation) gfil.ReverseExpansionDirectionOn() ls = gfil.Execute(init_ls, sitk.Cast(simg, sitk.sitkFloat32)) ls = label(sitk.GetArrayFromImage(ls) > 0) return keep_labels(labels, ls) def extract_large(labels, AREA=700): rps = regionprops(labels) retain = filter(lambda x:x.area>AREA, rps) badlabels = np.zeros(labels.shape, np.uint16) for r in retain: badlabels[labels==r.label] = r.label return badlabels def extract_small(labels, AREA=200): rps = regionprops(labels) retain = filter(lambda x:x.area>> q = [10, 10] >>> c = [0, 0] >>> p = [-10, 10] >>> calc_closewise_degree(p, c, q) 90.0 """ angle_r = cart2pol_angle(q[0]-c[0], q[1]-c[1]) - cart2pol_angle(p[0]-c[0], p[1]-c[1]) angle = 180.0 * angle_r/np.pi if angle < 0: angle += 360.0 return angle class CoordsConcave(object): """coords needs to be ordered in clockwise. """ def __init__(self, coords, edgelen=5, thres=180): self.coords = coords self.edgelen = edgelen self.thres = thres # def run(self): # score, coords = self.calc_neck_score_thres() # return coords def calc_neck_score(self): """Calculate the score (angle changes) and return the sorted score and the corresponding pixel coordinates. Pass the coordinates of outlines without border.""" ordered_c, edgelen = self.coords, self.edgelen nc = np.vstack((ordered_c, ordered_c[:edgelen, :])) score = [] for n, ci in enumerate(nc[:-edgelen, :]): try: score.append(calc_clockwise_degree(ordered_c[n-edgelen, :], nc[n, :], nc[n+edgelen, :])) except: pass idx = np.flipud(np.argsort(score)) return np.array(score)[idx], ordered_c[idx] def calc_neck_score_thres(self): score, s_coords = self.calc_neck_score() return score[score > self.thres], s_coords[score > self.thres] class CoordsConcaveWs(CoordsConcave): def __init__(self, coords, wlines, edgelen=5, thres=180): self.coords = coords self.edgelen = edgelen self.thres = thres self.wlines = wlines def run(self): score, coords = self.calc_neck_score_thres() return self.limit_coords_ws(coords), score def limit_coords_ws(self, coords): """modify later to find neighbors? """ cstore = [] for i in coords: if self.wlines[i[0], i[1]]: cstore.append(i) return np.array(cstore) def cellfilter(cell, small_area, large_area, major_minor=2.0): if cell.area > small_area and cell.area < large_area and cell.major_axis_length/cell.minor_axis_length < major_minor: return True else: return False class CellCutter(object): def __init__(self, bw, img, wlines, small_rad, large_rad=0, EDGELEN=10, THRES=180, CANDS_LIMIT=300): """ Use lower values for CANDS_LIMIT to speed up """ large_rad = 0 # fix it later. self.bw = bw self.img = img self.wlines = wlines self.EDGELEN = EDGELEN self.THRES = THRES self.filfunc = partial(cellfilter, small_area=np.pi*small_rad**2, large_area=np.pi*large_rad**2, major_minor=2.0) self.goodcells = [] # self.large_rad = large_rad self.small_rad = small_rad self.STEPLIM = self.small_rad * np.pi/2 self.coords_set = [] self.cut_coords = [] self.cut_cells = [] self.CANDS_LIMIT = CANDS_LIMIT def extract_cell_outlines(self): o_coords = find_oriented_coords(self.bw) if o_coords is not None: self.cell.o_coords = o_coords def run(self): self.prepare_coords_set() if not self.coords_set: return self.collect_goodcells() self.make_bw() if not self.cell: return while self.cell.area > self.large_rad ** 2 * np.pi: self.remove_coords_set() self.collect_goodcells() self.make_bw() if not self.cell: break def collect_goodcells(self): candidates = self.search_cut_candidates(self.bw.copy(), self.coords_set) if not candidates: return candidate = self.filter_candidates(candidates) if not candidate: return (n1, n2) = candidate.n1, candidate.n2 self.all_cells = make_candidates(self.bw.copy(), self.coords_set[n1][0][0], self.coords_set[n1][0][1], self.coords_set[n1][1][0], self.coords_set[n1][1][1]) self.cut_coords.append(self.coords_set[n1]) for c in self.all_cells: if self.filfunc(c): self.goodcells.append(c) else: self.cut_cells.append(c) def make_bw(self): if self.cut_cells: bw = np.zeros(self.bw.shape, np.uint8) for cell in self.cut_cells: for c in cell.coords: bw[c[0], c[1]] = 1 self.bw = bw self.cell = regionprops(bw, self.img)[0] self.cut_cells = [] else: self.cell = [] def remove_coords_set(self): store = [] for c0, c1 in self.coords_set: if self.bw[c0[0], c0[1]] and self.bw[c1[0], c1[1]]: store.append((c0, c1)) self.coords_set = store def prepare_coords_set(self): self.cell = regionprops(self.bw.astype(np.uint8), self.img)[0] if 1 in self.cell.image.shape: self.coords_set = [] return self.extract_cell_outlines() if not hasattr(self.cell, 'o_coords'): return coords, _ = CoordsConcaveWs(self.cell.o_coords, wlines=self.wlines, edgelen=self.EDGELEN, thres=self.THRES).run() if not coords.any(): return self.coords_set = self.make_sort_coordsset_by_dist(coords) if not self.coords_set: return self.coords_set = self.coords_set[:self.CANDS_LIMIT] self.coords_set = self.filter_coords_by_step(self.cell.o_coords, self.coords_set) def make_sort_coordsset_by_dist(self, coords): dist = cdist(coords, coords) boolarr = np.triu(np.ones(dist.shape, bool), 1) a1 = np.tile(range(len(coords)), len(coords)).reshape((len(coords), len(coords))) x1, x2 = a1[boolarr], a1.T[boolarr] sortorder = np.argsort(dist[boolarr]) x1, x2 = x1[sortorder], x2[sortorder] return [[coords[i1], coords[i2]] for i1, i2 in zip(x1, x2)] # def filter_coords_by_dist(self, coords): # dist = cdist(coords, coords) # boolarr = np.triu(dist < self.DISTLIM, 1) # a1 = np.tile(range(len(coords)), len(coords)).reshape((len(coords), len(coords))) # x1, x2 = a1[boolarr], a1.T[boolarr] # return [[coords[i1], coords[i2]] for i1, i2 in zip(x1, x2)] def filter_coords_by_step(self, coords, coords_set): new_coords_set = [] for c0, c1 in coords_set: min_step = calc_shortest_step_coords(coords, c0, c1) if min_step > self.STEPLIM: new_coords_set.append([c0, c1]) return new_coords_set def filter_candidates(self, candidates): candidates = filter(lambda x: x.area > np.pi * self.small_rad ** 2, candidates) if len(candidates): mindist = min([c.line_total for c in candidates]) index = [n for n, c in enumerate(candidates) if c.line_total == mindist][0] return candidates[index] else: return [] def search_cut_candidates(self, bw, coords_set): candidates = [] for n1, (startpt, endpt) in enumerate(coords_set): # cut and get candidate regionprops cand_rps = make_candidates_img(bw.copy(), startpt[0], startpt[1], endpt[0], endpt[1], self.img) if not len(cand_rps) > len(np.unique(label(self.cell.filled_image, connectivity=1))) - 1: continue if not all([c.area > np.pi * self.small_rad**2 for c in cand_rps]): continue # scoring for n2, c in enumerate(cand_rps): # If you get an error here, it might be unresolved error of scikit-image. https://github.com/scikit-image/scikit-image/issues/1470 # edit _regionprops._RegionProperties.moments and moments_central from "astype(np.uint8)" to "astype(np.float)" if c.minor_axis_length > 1: c.n1, c.n2 = n1, n2 c.cut_coords = (startpt, endpt) c.cutline_len = np.sqrt((startpt[0] - endpt[0])**2 + (startpt[1] - endpt[1])**2) c.line_total = intensity_below_line(self.img, startpt[0], startpt[1], endpt[0], endpt[1]).sum() candidates.append(c) return candidates def get_labels(self): temp = label(self.bw.copy(), connectivity=1) temp = np.zeros(self.bw.shape) if not self.goodcells: return label(self.bw) max_label = temp.max() for n, i in enumerate(self.all_cells): for c in i.coords: temp[c[0], c[1]] = max_label + n + 1 return temp def cut_labels(labels, coords_set): for i in coords_set: if i: labels = cut_neck(labels, i[0][0], i[0][1], i[1][0], i[1][1]) return labels def levelset_geo_separete(img, labels, niter=10, prop=-1, advec=-0.5, curve=-0.5): seg = sitk.GetImageFromArray((labels > 0).astype(np.uint8)) init_ls = sitk.SignedMaurerDistanceMap(seg, insideIsPositive=True, useImageSpacing=True) mask = find_boundaries(gvoronoi(label(labels, connectivity=1)), mode='inner') img1 = img.copy() img1[mask] = 0 img_T1 = sitk.GetImageFromArray(img1) lsFilter = sitk.GeodesicActiveContourLevelSetImageFilter() lsFilter.SetMaximumRMSError(0.02) lsFilter.SetNumberOfIterations(niter) lsFilter.SetAdvectionScaling(advec) lsFilter.SetCurvatureScaling(curve) lsFilter.SetPropagationScaling(prop) # lsFilter.ReverseExpansionDirectionOn() ls = lsFilter.Execute(init_ls, sitk.Cast(img_T1, sitk.sitkFloat32)) return label(sitk.GetArrayFromImage(ls) > 0, connectivity=1) def get_cut_coords(img, labels, small_rad, large_rad, EDGELEN=6, THRES=180): largela = extract_large(labels, AREA=np.pi * large_rad**2) wlines = wshed_raw(labels > 0, img) store = [] for i in np.unique(largela): if i == 0: continue cc = CellCutter(labels==i, img, wlines, small_rad=small_rad, large_rad=large_rad, EDGELEN=EDGELEN, THRES=THRES) cc.run() store.append(cc.cut_coords) return store def run_concave_cut(img, labels, small_rad, large_rad, EDGELEN=6, THRES=180): store = get_cut_coords(img, labels, small_rad, large_rad, EDGELEN=EDGELEN, THRES=THRES) labels = label(cut_labels(labels, [i for j in store for i in j]), connectivity=1) return labels PK!W$$celltk/utils/cp_functions.py""" Adapted from from CellProfiler """ import numpy as np import scipy.ndimage as scind import scipy.sparse from centrosome.filter import stretch from scipy.fftpack import fft2, ifft2 def align_cross_correlation(pixels1, pixels2): '''Align the second image with the first using max cross-correlation returns the x,y offsets to add to image1's indexes to align it with image2 Many of the ideas here are based on the paper, "Fast Normalized Cross-Correlation" by J.P. Lewis (http://www.idiom.com/~zilla/Papers/nvisionInterface/nip.html) which is frequently cited when addressing this problem. ''' # # TODO: Possibly use all 3 dimensions for color some day # if pixels1.ndim == 3: pixels1 = np.mean(pixels1, 2) if pixels2.ndim == 3: pixels2 = np.mean(pixels2, 2) # # We double the size of the image to get a field of zeros # for the parts of one image that don't overlap the displaced # second image. # # Since we're going into the frequency domain, if the images are of # different sizes, we can make the FFT shape large enough to capture # the period of the largest image - the smaller just will have zero # amplitude at that frequency. # s = np.maximum(pixels1.shape, pixels2.shape) fshape = s * 2 # # Calculate the # of pixels at a particular point # i, j = np.mgrid[-s[0]:s[0], -s[1]:s[1]] unit = np.abs(i * j).astype(float) unit[unit < 1] = 1 # keeps from dividing by zero in some places # # Normalize the pixel values around zero which does not affect the # correlation, keeps some of the sums of multiplications from # losing precision and precomputes t(x-u,y-v) - t_mean # pixels1 = pixels1 - np.mean(pixels1) pixels2 = pixels2 - np.mean(pixels2) # # Lewis uses an image, f and a template t. He derives a normalized # cross correlation, ncc(u,v) = # sum((f(x,y)-f_mean(u,v))*(t(x-u,y-v)-t_mean),x,y) / # sqrt(sum((f(x,y)-f_mean(u,v))**2,x,y) * (sum((t(x-u,y-v)-t_mean)**2,x,y) # # From here, he finds that the numerator term, f_mean(u,v)*(t...) is zero # leaving f(x,y)*(t(x-u,y-v)-t_mean) which is a convolution of f # by t-t_mean. # fp1 = fft2(pixels1, fshape) fp2 = fft2(pixels2, fshape) corr12 = ifft2(fp1 * fp2.conj()).real # # Use the trick of Lewis here - compute the cumulative sums # in a fashion that accounts for the parts that are off the # edge of the template. # # We do this in quadrants: # q0 q1 # q2 q3 # For the first, # q0 is the sum over pixels1[i:,j:] - sum i,j backwards # q1 is the sum over pixels1[i:,:j] - sum i backwards, j forwards # q2 is the sum over pixels1[:i,j:] - sum i forwards, j backwards # q3 is the sum over pixels1[:i,:j] - sum i,j forwards # # The second is done as above but reflected lr and ud # p1_si = pixels1.shape[0] p1_sj = pixels1.shape[1] p1_sum = np.zeros(fshape) p1_sum[:p1_si, :p1_sj] = cumsum_quadrant(pixels1, False, False) p1_sum[:p1_si, -p1_sj:] = cumsum_quadrant(pixels1, False, True) p1_sum[-p1_si:, :p1_sj] = cumsum_quadrant(pixels1, True, False) p1_sum[-p1_si:, -p1_sj:] = cumsum_quadrant(pixels1, True, True) # # Divide the sum over the # of elements summed-over # p1_mean = p1_sum / unit p2_si = pixels2.shape[0] p2_sj = pixels2.shape[1] p2_sum = np.zeros(fshape) p2_sum[:p2_si, :p2_sj] = cumsum_quadrant(pixels2, False, False) p2_sum[:p2_si, -p2_sj:] = cumsum_quadrant(pixels2, False, True) p2_sum[-p2_si:, :p2_sj] = cumsum_quadrant(pixels2, True, False) p2_sum[-p2_si:, -p2_sj:] = cumsum_quadrant(pixels2, True, True) p2_sum = np.fliplr(np.flipud(p2_sum)) p2_mean = p2_sum / unit # # Once we have the means for u,v, we can caluclate the # variance-like parts of the equation. We have to multiply # the mean^2 by the # of elements being summed-over # to account for the mean being summed that many times. # p1sd = np.sum(pixels1 ** 2) - p1_mean ** 2 * np.product(s) p2sd = np.sum(pixels2 ** 2) - p2_mean ** 2 * np.product(s) # # There's always chance of roundoff error for a zero value # resulting in a negative sd, so limit the sds here # sd = np.sqrt(np.maximum(p1sd * p2sd, 0)) corrnorm = corr12 / sd # # There's not much information for points where the standard # deviation is less than 1/100 of the maximum. We exclude these # from consideration. # corrnorm[(unit < np.product(s) / 2) & (sd < np.mean(sd) / 100)] = 0 i, j = np.unravel_index(np.argmax(corrnorm), fshape) # # Reflect values that fall into the second half # if i > pixels1.shape[0]: i = i - fshape[0] if j > pixels1.shape[1]: j = j - fshape[1] return j, i def align_mutual_information(pixels1, pixels2, mask1, mask2): '''Align the second image with the first using mutual information returns the x,y offsets to add to image1's indexes to align it with image2 The algorithm computes the mutual information content of the two images, offset by one in each direction (including diagonal) and then picks the direction in which there is the most mutual information. From there, it tries all offsets again and so on until it reaches a local maximum. ''' # # TODO: Possibly use all 3 dimensions for color some day # if pixels1.ndim == 3: pixels1 = np.mean(pixels1, 2) if pixels2.ndim == 3: pixels2 = np.mean(pixels2, 2) def mutualinf(x, y, maskx, masky): x = x[maskx & masky] y = y[maskx & masky] return entropy(x) + entropy(y) - entropy2(x, y) maxshape = np.maximum(pixels1.shape, pixels2.shape) pixels1 = reshape_image(pixels1, maxshape) pixels2 = reshape_image(pixels2, maxshape) mask1 = reshape_image(mask1, maxshape) mask2 = reshape_image(mask2, maxshape) best = mutualinf(pixels1, pixels2, mask1, mask2) i = 0 j = 0 while True: last_i = i last_j = j for new_i in range(last_i - 1, last_i + 2): for new_j in range(last_j - 1, last_j + 2): if new_i == 0 and new_j == 0: continue p2, p1 = offset_slice(pixels2, pixels1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) info = mutualinf(p1, p2, m1, m2) if info > best: best = info i = new_i j = new_j if i == last_i and j == last_j: return j, i, best def offset_slice(pixels1, pixels2, i, j): '''Return two sliced arrays where the first slice is offset by i,j relative to the second slice. ''' if i < 0: height = min(pixels1.shape[0] + i, pixels2.shape[0]) p1_imin = -i p2_imin = 0 else: height = min(pixels1.shape[0], pixels2.shape[0] - i) p1_imin = 0 p2_imin = i p1_imax = p1_imin + height p2_imax = p2_imin + height if j < 0: width = min(pixels1.shape[1] + j, pixels2.shape[1]) p1_jmin = -j p2_jmin = 0 else: width = min(pixels1.shape[1], pixels2.shape[1] - j) p1_jmin = 0 p2_jmin = j p1_jmax = p1_jmin + width p2_jmax = p2_jmin + width p1 = pixels1[p1_imin:p1_imax, p1_jmin:p1_jmax] p2 = pixels2[p2_imin:p2_imax, p2_jmin:p2_jmax] return p1, p2 def cumsum_quadrant(x, i_forwards, j_forwards): '''Return the cumulative sum going in the i, then j direction x - the matrix to be summed i_forwards - sum from 0 to end in the i direction if true j_forwards - sum from 0 to end in the j direction if true ''' if i_forwards: x = x.cumsum(0) else: x = np.flipud(np.flipud(x).cumsum(0)) if j_forwards: return x.cumsum(1) else: return np.fliplr(np.fliplr(x).cumsum(1)) def entropy(x): '''The entropy of x as if x is a probability distribution''' histogram = scind.histogram(x.astype(float), np.min(x), np.max(x), 256) n = np.sum(histogram) if n > 0 and np.max(histogram) > 0: histogram = histogram[histogram != 0] return np.log2(n) - np.sum(histogram * np.log2(histogram)) / n else: return 0 def entropy2(x, y): '''Joint entropy of paired samples X and Y''' # # Bin each image into 256 gray levels # x = (stretch(x) * 255).astype(int) y = (stretch(y) * 255).astype(int) # # create an image where each pixel with the same X & Y gets # the same value # xy = 256 * x + y xy = xy.flatten() sparse = scipy.sparse.coo_matrix((np.ones(xy.shape), (xy, np.zeros(xy.shape)))) histogram = sparse.toarray() n = np.sum(histogram) if n > 0 and np.max(histogram) > 0: histogram = histogram[histogram > 0] return np.log2(n) - np.sum(histogram * np.log2(histogram)) / n else: return 0 def reshape_image(source, new_shape): '''Reshape an image to a larger shape, padding with zeros''' if tuple(source.shape) == tuple(new_shape): return source result = np.zeros(new_shape, source.dtype) result[:source.shape[0], :source.shape[1]] = source return resultPK! 2ʤcelltk/utils/dlevel_set.pyimport numpy as np from subdetect_utils import calc_mask_exclude_overlap from filters import label def grad(x): return np.array(np.gradient(x)) def norm(x, axis=0): if x.ndim == 3: return np.sqrt(np.sum(np.square(x), axis=axis)) else: return np.abs(x) def div(fx, fy): fyy, fyx = grad(fy) fxy, fxx = grad(fx) return fxx + fyy def dot(x, y, axis=0): return np.sum(x * y, axis=axis) def dlevel_set(phi, F, niter=20, dt=-0.5, mask=None): """ Implementation of Level set method. https://wiseodd.github.io/techblog/2016/11/05/levelset-method/ It has an extra repulsion and masking processes. niter (int): iteration dt (float): negative values for propagation, positive values for shrinking phi (ndarray): labels, set inner objects as -1 and outside as 1. F (ndarray): img, set boundaries close to 0 and elsewhere close to 1. mask (ndarray[np.bool]): contours do not cross this line. """ if mask is None: mask = np.zeros(phi.shape, np.bool) for i in range(niter): dphi = grad(phi) dphi_norm = norm(dphi) region = calc_mask_exclude_overlap(label(phi < 0), 2) # added for absolute repulsion dphi_norm[region] = 0 # added for absolute repulsion dphi_norm[mask] = 0 # added constraints based on the image dphi_t = F * dphi_norm phi = phi + dt * dphi_t return phiPK!7]]celltk/utils/file_io.pyimport os import tifffile as tiff from os.path import basename, join import numpy as np import logging import tempfile import shutil import urllib logger = logging.getLogger(__name__) def make_dirs(path): try: os.makedirs(path) except OSError: if not os.path.isdir(path): raise def imsave(img, output, path, dtype=np.float32): """ save a single image or multiple images. len(path) == img.shape[2] """ if isinstance(path, list) or isinstance(path, tuple): for num, p in enumerate(path): filename = join(output, basename(p).split('.')[0]+'.tif') logger.debug('Image (shape {0}) is saved at {1}'.format(img.shape, filename)) if img.ndim == 2: tiff.imsave(filename, img.astype(dtype)) break tiff.imsave(filename, img[:, :, num].astype(dtype)) else: filename = join(output, basename(path).split('.')[0]+'.tif') logger.debug('Image (shape {0}) is saved at {1}'.format(img.shape, filename)) tiff.imsave(filename, img.astype(dtype)) def lbread(path, nonneg=True): def uint2int16(img): if (img == 0).any(): return img elif img.min() == 32768: return img - 32768 img = tiff.imread(path) img = uint2int16(img).astype(np.int16) if nonneg: img[img < 0] = 0 return img class LocalPath(object): """Generate a local path if URL is given. with LocalPath(path) as lpath: print(lpath) """ local = True def __init__(self, path): if os.path.exists(path): self.path = path else: self.local = False temp_dir = tempfile.mkdtemp() self.path = join(temp_dir, os.path.basename(path)) urllib.urlretrieve(path, self.path) def __enter__(self): return self.path def __exit__(self, *args): if not self.local: shutil.rmtree(os.path.dirname(self.path))PK!4`celltk/utils/filters.pyfrom __future__ import division import numpy as np from scipy.ndimage import distance_transform_edt from skimage.measure import regionprops from skimage.measure import label as skim_label from skimage.morphology import watershed as skiwatershed from skimage.feature import peak_local_max from skimage.segmentation import find_boundaries from skimage.feature import peak_local_max from scipy.ndimage.filters import maximum_filter from skimage.draw import line from scipy.ndimage.filters import gaussian_filter import SimpleITK as sitk from morphsnakes import MorphACWE, curvop from mahotas.segmentation import gvoronoi from skimage.morphology import thin import pandas as pd def label_watershed(labels, regmax): # Since there are non-unique values for dist, add very small numbers. This will separate each marker by regmax at least. dist = distance_transform_edt(labels) + np.random.rand(*labels.shape)*1e-10 labeled_maxima = label(peak_local_max(dist, min_distance=int(regmax), indices=False)) wshed = -dist wshed = wshed - np.min(dist) markers = np.zeros(wshed.shape, np.int16) markers[labeled_maxima > 0] = -labeled_maxima[labeled_maxima > 0] wlabel = skiwatershed(wshed, markers, connectivity=np.ones((3,3), bool), mask=labels!=0) wlabel = -wlabel wlabel = labels.max() + wlabel wlabel[wlabel == labels.max()] = 0 all_label = label(labels + wlabel) return all_label def label(bw, connectivity=2): '''original label might label any objects at top left as 1. To get around this pad it first.''' if bw[0, 0]: return skim_label(bw, connectivity=connectivity) bw = np.pad(bw, pad_width=1, mode='constant', constant_values=False) labels = skim_label(bw, connectivity=connectivity) labels = labels[1:-1, 1:-1] return labels def peak_local_max_edge(labels, min_dist=5): '''peak_local_max sometimes shows a weird behavior...?''' label_max = maximum_filter(labels, size=min_dist) mask = label == label_max label[-mask] = 0 return labels def find_label_boundaries(labels): blabels = labels.copy() bwbound = find_boundaries(blabels) blabels[-bwbound] = 0 return blabels def labels2outlines(labels): """Same functionality with find_label_boundaries. """ outlines = labels.copy() outlines[~find_boundaries(labels)] = 0 return outlines def adaptive_thresh(img, R=1, FILTERINGSIZE=50): """Segment as a foreground if pixel is higher than ratio * blurred image. If you set R=10, it will pick a pixel if a pixel in the raw image is at least 10% brighter than the blurred image. """ fim = gaussian_filter(img, FILTERINGSIZE) bw = img > (fim * (1. + R/100.)) return bw def calc_lapgauss(img, SIGMA=2.5): fil = sitk.LaplacianRecursiveGaussianImageFilter() fil.SetSigma(SIGMA) # fil.SetNormalizeAcrossScale(False) csimg = sitk.GetImageFromArray(img) slap = fil.Execute(csimg) return sitk.GetArrayFromImage(slap) def gray_fill_holes(labels): '''This will fill holes of gray int images''' # labels = np.int32(labels) # labels = np.pad(labels, pad_width=1, mode='constant', constant_values=-100000) # blabel = labels.copy() # blabel[1:-1, 1:-1] = 100000 # fim = reconstruction(neg(blabel), neg(labels)) # fim = neg(np.int32(fim)) # fim = fim[1:-1, 1:-1] # return fim fil = sitk.GrayscaleFillholeImageFilter() return sitk.GetArrayFromImage(fil.Execute(sitk.GetImageFromArray(labels))) def sitk_watershed_intensity(img, local_maxima): seedimage = sitk.GetImageFromArray(local_maxima.astype(np.uint16))# img = img.astype(np.float32) nimg = sitk.GetImageFromArray(img) nimg = sitk.GradientMagnitude(nimg)# fil = sitk.MorphologicalWatershedFromMarkersImageFilter() fil.FullyConnectedOn() fil.MarkWatershedLineOff() oimg1 = fil.Execute(nimg, seedimage) labelim = sitk.GetArrayFromImage(oimg1) return labelim def lap_local_max(img, sigma_list, THRES): img = np.uint16(img) lapimages = [] for sig in sigma_list: simg = sitk.GetImageFromArray(img) nimg = sitk.LaplacianRecursiveGaussian(image1=simg, sigma=sig) lapimages.append(-sitk.GetArrayFromImage(nimg)) image_cube = np.dstack(lapimages) local_maxima = peak_local_max(image_cube, threshold_abs=THRES, footprint=np.ones((3, 3, 3)), threshold_rel=0.0, exclude_border=False, indices=False) local_maxima = local_maxima.sum(axis=2) local_maxima = label(local_maxima) return local_maxima class MultiSnakes(MorphACWE): def __init__(self, img, labels, smoothing=1, lambda1=1, lambda2=1): super(MultiSnakes, self).__init__(img, smoothing, lambda1, lambda2) self.levelset = labels def multi_step(self, niter=1): for i in range(niter): self.step() return self.return_labels() def step(self): # Assign attributes to local variables for convenience. u = self._u if u is None: raise ValueError("the levelset function is not set (use set_levelset)") data = self.data # Determine c0 and c1. inside = u>0 outside = u<=0 c0 = data[outside].sum() / float(outside.sum()) c1 = data[inside].sum() / float(inside.sum()) # Image attachment. dres = np.array(np.gradient(u)) abs_dres = np.abs(dres).sum(0) aux = abs_dres * (self.lambda1*(data - c1)**2 - self.lambda2*(data - c0)**2) mask = find_boundaries(gvoronoi(label(u, connectivity=1)), mode='inner') aux[mask] = 1 res = np.copy(u) res[aux < 0] = 1 res[aux > 0] = 0 # Smoothing. for i in range(self.smoothing): res = curvop(res) self._u = res def return_labels(self): return label(self.levelset, connectivity=1) class MultiSnakesCombined(MultiSnakes): def multi_step(self, niter=1): for i in range(niter-1): self.step() self.step_last() return self.return_labels() def step_last(self): # Assign attributes to local variables for convenience. u = self._u mask = thin(find_boundaries(gvoronoi(label(u, connectivity=1)), mode='inner')) if u is None: raise ValueError("the levelset function is not set (use set_levelset)") data = self.data # Determine c0 and c1. inside = u>0 outside = u<=0 c0 = data[outside].sum() / float(outside.sum()) c1 = data[inside].sum() / float(inside.sum()) # Image attachment. dres = np.array(np.gradient(u)) abs_dres = np.abs(dres).sum(0) aux = abs_dres * (self.lambda1*(data - c1)**2 - self.lambda2*(data - c0)**2) res = np.copy(u) res[aux < 0] = 1 res[aux > 0] = 0 # Smoothing. for i in range(self.smoothing): res = curvop(res) res[mask] = 0 self._u = res def interpolate_nan(arr): """Approximate a linear interpolation of array with NaNs. """ arr[arr < 0] = np.nan h_interp = pd.DataFrame(arr).interpolate(axis=0) w_interp = pd.DataFrame(arr).interpolate(axis=1) interpolated = np.nanmean(np.dstack((h_interp, w_interp)), axis=2) interpolated[np.isnan(interpolated)] = 0 return interpolated PK!2GYcelltk/utils/fish_detect.pyimport numpy as np import pywt from skimage.transform import resize from skimage.measure import label from filters import adaptive_thresh def enhance_puncta(img, level=7): """ Removing low frequency wavelet signals to enhance puncta. Dependent on image size, try level 6~8. """ if level == 0: return img wp = pywt.WaveletPacket2D(data=img, wavelet='haar', mode='sym') back = resize(np.array(wp['d'*level].data), img.shape, order=3, mode='reflect')/(2**level) cimg = img - back cimg[cimg < 0] = 0 return cimg def detect_puncta(img, level=7, PERC=50, FILSIZE=1): pimg = enhance_puncta(img.astype(np.uint16), level) limg = label(adaptive_thresh(pimg, R=PERC, FILTERINGSIZE=FILSIZE), connectivity=1) return limgPK!M:celltk/utils/global_holder.py def counter(): i = 0 while True: i += 1 yield i class Holder(object): c = counter() def count(self): return self.c.next() # global holder holder = Holder() PK!jf$$celltk/utils/imreg.py# -*- coding: utf-8 -*- # -*- coding: utf-8 -*- # imreg.py # Copyright (c) 2011-2014, Christoph Gohlke # Copyright (c) 2011-2014, The Regents of the University of California # Produced at the Laboratory for Fluorescence Dynamics # All rights reserved. # # Redistribution and use in source and binary forms, with or without # modification, are permitted provided that the following conditions are met: # # * Redistributions of source code must retain the above copyright # notice, this list of conditions and the following disclaimer. # * Redistributions in binary form must reproduce the above copyright # notice, this list of conditions and the following disclaimer in the # documentation and/or other materials provided with the distribution. # * Neither the name of the copyright holders nor the names of any # contributors may be used to endorse or promote products derived # from this software without specific prior written permission. # # THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS" # AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE # IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE # ARE DISCLAIMED. IN NO EVENT SHALL THE COPYRIGHT OWNER OR CONTRIBUTORS BE # LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR # CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF # SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS # INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN # CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) # ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE # POSSIBILITY OF SUCH DAMAGE. """FFT based image registration. Implements an FFT-based technique for translation, rotation and scale-invariant image registration [1]. :Author: `Christoph Gohlke `_ :Organization: Laboratory for Fluorescence Dynamics, University of California, Irvine :Version: 2013.01.18 Requirements ------------ * `CPython 2.7 or 3.3 `_ * `Numpy 1.7 `_ * `Scipy 0.12 `_ * `Matplotlib 1.2 `_ (optional for plotting) Notes ----- The API and algorithms are not stable yet and are expected to change between revisions. References ---------- (1) An FFT-based technique for translation, rotation and scale-invariant image registration. BS Reddy, BN Chatterji. IEEE Transactions on Image Processing, 5, 1266-1271, 1996 (2) An IDL/ENVI implementation of the FFT-based algorithm for automatic image registration. H Xiea, N Hicksa, GR Kellera, H Huangb, V Kreinovich. Computers & Geosciences, 29, 1045-1055, 2003. (3) Image Registration Using Adaptive Polar Transform. R Matungka, YF Zheng, RL Ewing. IEEE Transactions on Image Processing, 18(10), 2009. Examples -------- >>> im0 = imread('t400') >>> im1 = imread('Tr19s1.3') >>> im2, scale, angle, (t0, t1) = similarity(im0, im1) >>> imshow(im0, im1, im2) >>> im0 = imread('t350380ori') >>> im1 = imread('t350380shf') >>> t0, t1 = translation(im0, im1) """ from __future__ import division, print_function import math import numpy from numpy.fft import fft2, ifft2, fftshift try: import scipy.ndimage.interpolation as ndii except ImportError: import ndimage.interpolation as ndii __version__ = '2013.01.18' __docformat__ = 'restructuredtext en' __all__ = ['translation', 'similarity'] def translation(im0, im1): """Return translation vector to register images.""" shape = im0.shape f0 = fft2(im0) f1 = fft2(im1) ir = abs(ifft2((f0 * f1.conjugate()) / (abs(f0) * abs(f1)))) t0, t1 = numpy.unravel_index(numpy.argmax(ir), shape) if t0 > shape[0] // 2: t0 -= shape[0] if t1 > shape[1] // 2: t1 -= shape[1] return [t0, t1] def similarity(im0, im1): """Return similarity transformed image im1 and transformation parameters. Transformation parameters are: isotropic scale factor, rotation angle (in degrees), and translation vector. A similarity transformation is an affine transformation with isotropic scale and without shear. Limitations: Image shapes must be equal and square. All image areas must have same scale, rotation, and shift. Scale change must be less than 1.8. No subpixel precision. """ if im0.shape != im1.shape: raise ValueError("Images must have same shapes.") elif len(im0.shape) != 2: raise ValueError("Images must be 2 dimensional.") f0 = fftshift(abs(fft2(im0))) f1 = fftshift(abs(fft2(im1))) h = highpass(f0.shape) f0 *= h f1 *= h del h f0, log_base = logpolar(f0) f1, log_base = logpolar(f1) f0 = fft2(f0) f1 = fft2(f1) r0 = abs(f0) * abs(f1) ir = abs(ifft2((f0 * f1.conjugate()) / r0)) i0, i1 = numpy.unravel_index(numpy.argmax(ir), ir.shape) angle = 180.0 * i0 / ir.shape[0] scale = log_base ** i1 if scale > 1.8: ir = abs(ifft2((f1 * f0.conjugate()) / r0)) i0, i1 = numpy.unravel_index(numpy.argmax(ir), ir.shape) angle = -180.0 * i0 / ir.shape[0] scale = 1.0 / (log_base ** i1) if scale > 1.8: raise ValueError("Images are not compatible. Scale change > 1.8") if angle < -90.0: angle += 180.0 elif angle > 90.0: angle -= 180.0 im2 = ndii.zoom(im1, 1.0/scale) im2 = ndii.rotate(im2, angle) if im2.shape < im0.shape: t = numpy.zeros_like(im0) t[:im2.shape[0], :im2.shape[1]] = im2 im2 = t elif im2.shape > im0.shape: im2 = im2[:im0.shape[0], :im0.shape[1]] f0 = fft2(im0) f1 = fft2(im2) ir = abs(ifft2((f0 * f1.conjugate()) / (abs(f0) * abs(f1)))) t0, t1 = numpy.unravel_index(numpy.argmax(ir), ir.shape) if t0 > f0.shape[0] // 2: t0 -= f0.shape[0] if t1 > f0.shape[1] // 2: t1 -= f0.shape[1] im2 = ndii.shift(im2, [t0, t1]) # correct parameters for ndimage's internal processing if angle > 0.0: d = int((int(im1.shape[1] / scale) * math.sin(math.radians(angle)))) t0, t1 = t1, d+t0 elif angle < 0.0: d = int((int(im1.shape[0] / scale) * math.sin(math.radians(angle)))) t0, t1 = d+t1, d+t0 scale = (im1.shape[1] - 1) / (int(im1.shape[1] / scale) - 1) return im2, scale, angle, [-t0, -t1] def similarity_matrix(scale, angle, vector): """Return homogeneous transformation matrix from similarity parameters. Transformation parameters are: isotropic scale factor, rotation angle (in degrees), and translation vector (of size 2). The order of transformations is: scale, rotate, translate. """ S = numpy.diag([scale, scale, 1.0]) R = numpy.identity(3) angle = math.radians(angle) R[0, 0] = math.cos(angle) R[1, 1] = math.cos(angle) R[0, 1] = -math.sin(angle) R[1, 0] = math.sin(angle) T = numpy.identity(3) T[:2, 2] = vector return numpy.dot(T, numpy.dot(R, S)) def logpolar(image, angles=None, radii=None): """Return log-polar transformed image and log base.""" shape = image.shape center = shape[0] / 2, shape[1] / 2 if angles is None: angles = shape[0] if radii is None: radii = shape[1] theta = numpy.empty((angles, radii), dtype=numpy.float64) theta.T[:] = -numpy.linspace(0, numpy.pi, angles, endpoint=False) #d = radii d = numpy.hypot(shape[0]-center[0], shape[1]-center[1]) log_base = 10.0 ** (math.log10(d) / (radii)) radius = numpy.empty_like(theta) radius[:] = numpy.power(log_base, numpy.arange(radii, dtype=numpy.float64)) - 1.0 x = radius * numpy.sin(theta) + center[0] y = radius * numpy.cos(theta) + center[1] output = numpy.empty_like(x) ndii.map_coordinates(image, [x, y], output=output) return output, log_base def highpass(shape): """Return highpass filter to be multiplied with fourier transform.""" x = numpy.outer( numpy.cos(numpy.linspace(-math.pi/2., math.pi/2., shape[0])), numpy.cos(numpy.linspace(-math.pi/2., math.pi/2., shape[1]))) return (1.0 - x) * (2.0 - x) def imread(fname, norm=True): """Return image data from img&hdr uint8 files.""" with open(fname+'.hdr', 'r') as fh: hdr = fh.readlines() img = numpy.fromfile(fname+'.img', numpy.uint8, -1) img.shape = int(hdr[4].split()[-1]), int(hdr[3].split()[-1]) if norm: img = img.astype(numpy.float64) img /= 255.0 return img def imshow(im0, im1, im2, im3=None, cmap=None, **kwargs): """Plot images using matplotlib.""" from matplotlib import pyplot if cmap is None: cmap = 'coolwarm' if im3 is None: im3 = abs(im2 - im0) pyplot.subplot(221) pyplot.imshow(im0, cmap, **kwargs) pyplot.subplot(222) pyplot.imshow(im1, cmap, **kwargs) pyplot.subplot(223) pyplot.imshow(im3, cmap, **kwargs) pyplot.subplot(224) pyplot.imshow(im2, cmap, **kwargs) pyplot.show()PK!we..celltk/utils/labels_handling.pyfrom filters import label import numpy as np def labels_map(lb0, lb1): """lb0 and lb1 should have objects in the same locations but different labels. """ lbnums = np.unique(lb0).tolist() lbnums.remove(0) st = [] for n0 in lbnums: n_sets = set(lb1[lb0 == n0]) assert not len(n_sets) == 0 n1 = list(n_sets)[0] st.append((n0, n1)) return st def convert_labels(lb_ref_to, lb_ref_from, lb_convert): """ Maps a reference set of labels onto another set of labels for the same image Example: Separate segmentation of two objects in an image (nucleus and cytoplasm) leads to different labels and can reconcile the same cell to have the same label for both objects. Args: lb_ref_to (np.ndarray): image with object labels to convert to lb_ref_from (np.ndarray): image with object labels to be converted lb_convert (np.ndarray): the labeled image to be converted Returns: arr (numpy.ndarray): converted labels for objects with reference set of labels """ lbmap_to, lbmap_from = zip(*labels_map(lb_ref_to, lb_ref_from)) arr = np.zeros(lb_convert.shape, dtype=np.uint16) lb = lb_convert.copy() for n0, n1 in zip(lbmap_to, lbmap_from): arr[lb == n0] = n1 return arr PK!JJcelltk/utils/mi_align.py""" Image alignment based on mutual information. Some functions are adapted/modified from cellprofiler. """ from __future__ import division import scipy.ndimage as scind import numpy as np import scipy from skimage.measure import block_reduce from scipy.ndimage.filters import gaussian_laplace from skimage.exposure import equalize_adapthist, equalize_hist from imreg import translation from scipy.ndimage import imread # from numba.decorators import jit from centrosome.filter import stretch # @jit # def stretch(image): # '''Normalize an image to make the minimum zero and maximum one # image - pixel data to be normalized # mask - optional mask of relevant pixels. None = don't mask # returns the stretched image # ''' # return (image - image.min()) / (image.max() - image.min()) class BaseMutualInfoAligner(object): def __init__(self, img1, img2, mask0=None, DOWNSAMPLE=(16, 8, 4, 2)): self.img1, self.img2 = img1, img2 if mask0 is None: mask0 = np.ones(img1.shape, bool) self.mask0 = mask0 self.DOWNSAMPLE = DOWNSAMPLE def execute(self): self.preprocessing() self.initial_mi() self.mi_loop() self.mi_last() def preprocessing(self): self.img1, self.img2 = equalize_adapthist(self.img1), equalize_adapthist(self.img2) def initial_mi(self): dimg1, dimg2, dmask = self.downsampling(self.DOWNSAMPLE[0]) arr, i_list, j_list = self.search_all_mi(dimg1, dimg2, dmask, dmask) self.ini_j, self.ini_i = self.pick_coords(arr, i_list, j_list) def downsampling(self, factor): # [1:-1, 1:-1] added in case a image has odd shape. dimg1 = block_reduce(self.img1, (factor, factor))[1:-1, 1:-1] dimg2 = block_reduce(self.img2, (factor, factor))[1:-1, 1:-1] dmask = block_reduce(self.mask0, (factor, factor))[1:-1, 1:-1] dmask = dmask!=0 return dimg1, dimg2, dmask def search_all_mi(self, pp1, pp2, mask1, mask2): best = 0 i_list = range(-pp1.shape[0], pp1.shape[0]) j_list = range(-pp1.shape[1], pp1.shape[1]) arr = np.zeros((len(i_list), len(j_list))) for n1, new_i in enumerate(i_list): for n2, new_j in enumerate(j_list): p2, p1 = offset_slice(pp2, pp1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) if p2.any() and p1.any() and m1.any() and m2.any(): info = mutualinf(p1, p2, m1, m2) info = info * p1.size/pp1.size # Prevent images to be aligned at the edge. Weight more on the center. arr[n1, n2] = info self.arr, self.i_list, self.j_list = arr, i_list, j_list return arr, i_list, j_list def pick_coords(self, arr, i_list, j_list): a1, a2 = np.where(arr == arr.max()) return j_list[a2], i_list[a1] def mi_loop(self): pass def mi_last(self): pass class BaseMutualInfoAlignerLog(BaseMutualInfoAligner): def search_all_mi(self, pp1, pp2, mask1, mask2): CUT = 10 best = 0 i_list = range(-pp1.shape[0]+CUT, pp1.shape[0]-CUT) j_list = range(-pp1.shape[1]+CUT, pp1.shape[1]-CUT) arr = np.zeros((len(i_list), len(j_list))) for n1, new_i in enumerate(i_list): for n2, new_j in enumerate(j_list): p2, p1 = offset_slice(pp2, pp1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) if p2.any() and p1.any() and m1.any() and m2.any(): info = mutualinf(p1, p2, m1, m2) info = info * p1.size/pp1.size # Prevent images to be aligned at the edge. Weight more on the center. arr[n1, n2] = info self.arr, self.i_list, self.j_list = arr, i_list, j_list return arr, i_list, j_list def pick_coords(self, arr, i_list, j_list): self.gl = gaussian_laplace(arr, 1) a1, a2 = np.where(self.gl==self.gl.min()) a1, a2 = a1[0], a2[0] return j_list[a2], i_list[a1] class MutualInfoAlignerLogLoop(BaseMutualInfoAlignerLog): def mi_loop(self): tj, ti = self.ini_j, self.ini_i for num, di in enumerate(self.DOWNSAMPLE[1:]): dimg1, dimg2, dmask = self.downsampling(di) tj, ti = tj*int(self.DOWNSAMPLE[num]/di), ti*int(self.DOWNSAMPLE[num]/di) best = mutualinf(dimg1, dimg2, dmask, dmask) tj, ti, _ = self.optimize_max_mi_from_offset(dimg1, dimg2, dmask, dmask, ti, tj, best) self._j, self._i = tj, ti def mi_last(self): tj, ti = self._j * self.DOWNSAMPLE[-1], self._i * self.DOWNSAMPLE[-1] best = mutualinf(self.img1, self.img2, self.mask0, self.mask0) tj, ti, mi = self.optimize_max_mi_from_offset(self.img1, self.img2, self.mask0, self.mask0, ti, tj, best) self._j, self._i, self.mi = tj, ti, mi def optimize_max_mi_from_offset(self, pixels1, pixels2, mask1, mask2, i, j, best): while True: last_i = i last_j = j for new_i in range(last_i - 1, last_i + 2): for new_j in range(last_j - 1, last_j + 2): if new_i == 0 and new_j == 0: continue p2, p1 = offset_slice(pixels2, pixels1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) if p1[m1].any() and p2[m2].any(): info = mutualinf(p1, p2, m1, m2) info = info * p1.size/pixels1.size # Prevent images to be aligned at the edge. Weight more on the center. else: info = 0 if info > best: best = info i = new_i j = new_j if i == last_i and j == last_j: return j, i, best class MutualInfoAlignerMultiHypothesis(MutualInfoAlignerLogLoop): """ Calculate all the possible MI in the downsampled image and then search local MI in a less downsampled image based. """ def execute(self): self.preprocessing() self.initial_mi() self.ini_j_all = self.ini_j[:] self.ini_i_all = self.ini_i[:] store, mistore = [], [] for self.ini_j, self.ini_i in zip(self.ini_j_all, self.ini_i_all): self.mi_loop() self.mi_last() store.append((self._j, self._i)) mistore.append(self.mi) j, i = store[mistore.index(max(mistore))] self.store = store self.mistore = mistore self._j, self._i, self.mi = j, i, max(mistore) def pick_coords(self, arr, i_list, j_list): HYPNUM = 5 gl = gaussian_laplace(arr, 1) # Divide images into 25 blocks and calculate minimum BLK = 5 hbl, wbl = np.linspace(0, gl.shape[0], BLK, dtype=int), np.linspace(0, gl.shape[1], BLK, dtype=int) gl_min = [] for (h1, h2) in zip(hbl[:-1], hbl[1:]): for (w1, w2) in zip(wbl[:-1], wbl[1:]): gl_min.append(gl[h1:h2, w1:w2].min()) gl_min.sort() ji_list = [] # plt.imshow(arr) # Pick top HYPNUM minimum from 25 blocks. j_all, i_all = [], [] for ii in range(HYPNUM): a1, a2 = np.where(gl==gl_min[ii]) # if len(a1) > 1: a1, a2 = a1[0], a2[0] j_all.append(j_list[a2]) i_all.append(i_list[a1]) return j_all, i_all def mutualinf(x, y, maskx, masky): x = x[maskx & masky] y = y[maskx & masky] return entropy(x) + entropy(y) - entropy2(x, y) def crop_images_based_on_r1_r2(img1, img2, r1, r2): if r2<0 and r1<0: cimg1, cimg2 = img1[:r2, :r1], img2[-r2:, -r1:] elif r2<0 and r1>0: cimg1, cimg2 = img1[:r2, r1:], img2[-r2:, :-r1] elif r2>0 and r1<0: cimg1, cimg2 = img1[r2:, :r1], img2[:-r2, -r1:] elif r2>0 and r1>0: cimg1, cimg2 = img1[r2:, r1:], img2[:-r2, :-r1] return cimg1, cimg2 def mask_image_edge(img, wpix, hpix): mask = np.ones(img.shape, bool) mask[hpix:-hpix, wpix:-wpix] = False return mask def align_mutual_information(pixels1, pixels2, mask1, mask2): '''Align the second image with the first using mutual information returns the x,y offsets to add to image1's indexes to align it with image2 The algorithm computes the mutual information content of the two images, offset by one in each direction (including diagonal) and then picks the direction in which there is the most mutual information. From there, it tries all offsets again and so on until it reaches a local maximum. ''' # # TODO: Possibly use all 3 dimensions for color some day # if pixels1.ndim == 3: pixels1 = np.mean(pixels1, 2) if pixels2.ndim == 3: pixels2 = np.mean(pixels2, 2) def mutualinf(x, y, maskx, masky): x = x[maskx & masky] y = y[maskx & masky] return entropy(x) + entropy(y) - entropy2(x, y) maxshape = np.maximum(pixels1.shape, pixels2.shape) pixels1 = reshape_image(pixels1, maxshape) pixels2 = reshape_image(pixels2, maxshape) mask1 = reshape_image(mask1, maxshape) mask2 = reshape_image(mask2, maxshape) best = mutualinf(pixels1, pixels2, mask1, mask2) i = 0 j = 0 while True: last_i = i last_j = j for new_i in range(last_i - 1, last_i + 2): for new_j in range(last_j - 1, last_j + 2): if new_i == 0 and new_j == 0: continue p2, p1 = offset_slice(pixels2, pixels1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) info = mutualinf(p1, p2, m1, m2) if info > best: best = info i = new_i j = new_j if i == last_i and j == last_j: return j, i, best def offset_slice(pixels1, pixels2, i, j): '''Return two sliced arrays where the first slice is offset by i,j relative to the second slice. ''' if i < 0: height = min(pixels1.shape[0] + i, pixels2.shape[0]) p1_imin = -i p2_imin = 0 else: height = min(pixels1.shape[0], pixels2.shape[0] - i) p1_imin = 0 p2_imin = i p1_imax = p1_imin + height p2_imax = p2_imin + height if j < 0: width = min(pixels1.shape[1] + j, pixels2.shape[1]) p1_jmin = -j p2_jmin = 0 else: width = min(pixels1.shape[1], pixels2.shape[1] - j) p1_jmin = 0 p2_jmin = j p1_jmax = p1_jmin + width p2_jmax = p2_jmin + width p1 = pixels1[p1_imin:p1_imax, p1_jmin:p1_jmax] p2 = pixels2[p2_imin:p2_imax, p2_jmin:p2_jmax] return p1, p2 # @jit def call_hist(x): _bins = np.linspace(x.min(), x.max(), 257) return np.histogram(x, _bins)[0] def entropy(x): '''The entropy of x as if x is a probability distribution''' # histogram = scind.histogram(x.astype(float), np.min(x), np.max(x), 256) histogram = call_hist(x) n = np.sum(histogram) if n > 0 and np.max(histogram) > 0: histogram = histogram[histogram != 0] return np.log2(n) - np.sum(histogram * np.log2(histogram)) / n else: return 0 def entropy2(x, y): '''Joint entropy of paired samples X and Y''' # # Bin each image into 256 gray levels # x = (stretch(x) * 255).astype(int) y = (stretch(y) * 255).astype(int) # # create an image where each pixel with the same X & Y gets # the same value # xy = 256 * x + y xy = xy.flatten() sparse = scipy.sparse.coo_matrix((np.ones(xy.shape), (xy, np.zeros(xy.shape)))) histogram = sparse.toarray() n = np.sum(histogram) if n > 0 and np.max(histogram) > 0: histogram = histogram[histogram > 0] return np.log2(n) - np.sum(histogram * np.log2(histogram)) / n else: return 0 def reshape_image(source, new_shape): '''Reshape an image to a larger shape, padding with zeros''' if tuple(source.shape) == tuple(new_shape): return source result = np.zeros(new_shape, source.dtype) result[:source.shape[0], :source.shape[1]] = source return result def calc_jitters(img0, img1): """ Take two images and calculate jitters. compare it to FFT based alignment and pick ones with higher mutual info. align_mutual_information can often stack in a local solution. """ mask = np.ones(img0.shape, np.bool) j0, j1, score0 = align_mutual_information(img0, img1, mask, mask) jj1, jj0 = translation(img0, img1) p2, p1 = offset_slice(img1, img0, jj1, jj0) m2, m1 = offset_slice(mask, mask, jj1, jj0) score1 = mutualinf(p1, p2, m1, m2) if score1 > score0: j0, j1 = jj0, jj1 return j0, j1 def calc_jitters_multiple(inputs, ch=100, cw=100): """ Calculate how many pixels images move based on the first image coordinates. Take multiple images and calculate jitters for consecutive images. INPUT: inputs(List): path to images """ jitters = [(0, 0)] for path0, path1 in zip(inputs[:-1], inputs[1:]): img0 = imread(path0)[ch:-ch, cw:-cw] img1 = imread(path1)[ch:-ch, cw:-cw] j0, j1 = calc_jitters(img0, img1) jitters.append((jitters[-1][0] + j0, jitters[-1][1] + j1)) return jitters def calc_crop_coordinates(store, shapes): """ Calculate how each images should be cropped. Take output from calc_jitters_multiple and return (start, end) coordinates for x, y for each image. """ max_w = max(i[0] for i in store) start_w = [max_w - i[0] for i in store] size_w = min([shapes[1] + i[0] for i in store]) - max_w max_h = max(i[1] for i in store) start_h = [max_h - i[1] for i in store] size_h = min([shapes[0] + i[1] for i in store]) - max_h return [(hi, hi+size_h, wi, wi+size_w) for hi, wi in zip(start_h, start_w)] class MinimalMutualInfoAligner(object): """ Use downsample and a part of image to calculate mutual information based jitters in a faster manner. """ def __init__(self, img1, img2, MAXIMUM=240, DOWNSAMPLE=4, preprocess=False): self.img1, self.img2 = img1, img2 self.mask0 = np.ones(img1.shape, bool) self.DOWNSAMPLE = DOWNSAMPLE self.MAXIMUM = MAXIMUM self.preprocess = preprocess def preprocessing(self): if self.preprocess: self.img1, self.img2 = equalize_hist(self.img1), equalize_hist(self.img2) def initial_mi(self): dimg1, dimg2, dmask = self.downsampling(self.DOWNSAMPLE) self.ini_j, self.ini_i = [], [] size = int(self.MAXIMUM/self.DOWNSAMPLE) TRIAL = 3 for i in range(TRIAL): hstart = np.random.randint(dimg1.shape[0] - size) wstart = np.random.randint(dimg1.shape[1] - size) ddimg1 = dimg1[hstart:hstart+size, wstart:wstart+size] ddimg2 = dimg2[hstart:hstart+size, wstart:wstart+size] ddmask = dmask[hstart:hstart+size, wstart:wstart+size] arr, i_list, j_list = self.search_all_mi(ddimg1, ddimg2, ddmask, ddmask) j0, i0 = self.pick_coords(arr, i_list, j_list) self.ini_j.append(j0) self.ini_i.append(i0) def downsampling(self, factor): # [1:-1, 1:-1] added in case a image has odd shape. dimg1 = block_reduce(self.img1, (factor, factor))[1:-1, 1:-1] dimg2 = block_reduce(self.img2, (factor, factor))[1:-1, 1:-1] dmask = block_reduce(self.mask0, (factor, factor))[1:-1, 1:-1] dmask = dmask != 0 return dimg1, dimg2, dmask def pick_coords(self, arr, i_list, j_list): # cut 10% due to the instability of GL at edges cri, crj = int(arr.shape[0] * 0.1), int(arr.shape[1] * 0.1) arr = arr[cri:-cri, crj:-crj] i_list, j_list = i_list[cri:-cri], j_list[crj:-crj] self.gl = gaussian_laplace(arr, 1) a1, a2 = np.where(self.gl == self.gl.min()) return j_list[a2[0]], i_list[a1[0]] def mi_last(self): self._j, self._i = self.ini_j, self.ini_i tj, ti = self._j * self.DOWNSAMPLE, self._i * self.DOWNSAMPLE best = mutualinf(self.img1, self.img2, self.mask0, self.mask0) tj, ti, mi = self.optimize_max_mi_from_offset(self.img1, self.img2, self.mask0, self.mask0, ti, tj, best) self._j, self._i, self.mi = tj, ti, mi def optimize_max_mi_from_offset(self, pixels1, pixels2, mask1, mask2, i, j, best): while True: last_i = i last_j = j for new_i in range(last_i - 1, last_i + 2): for new_j in range(last_j - 1, last_j + 2): if new_i == 0 and new_j == 0: continue p2, p1 = offset_slice(pixels2, pixels1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) if p1[m1].any() and p2[m2].any(): info = mutualinf(p1, p2, m1, m2) else: info = 0 if info > best: best = info i = new_i j = new_j if i == last_i and j == last_j: return j, i, best def search_all_mi(self, pp1, pp2, mask1, mask2): i_list = range(-pp1.shape[0], pp1.shape[0]) j_list = range(-pp1.shape[1], pp1.shape[1]) arr = np.zeros((len(i_list), len(j_list))) for n1, new_i in enumerate(i_list): for n2, new_j in enumerate(j_list): p2, p1 = offset_slice(pp2, pp1, new_i, new_j) m2, m1 = offset_slice(mask2, mask1, new_i, new_j) if p2.any() and p1.any() and m1.any() and m2.any(): info = mutualinf(p1, p2, m1, m2) arr[n1, n2] = info * (m1.shape[0] + m1.shape[1])/2 # add bias to the middle self.arr, self.i_list, self.j_list = arr, i_list, j_list return arr, i_list, j_list def execute(self): self.preprocessing() self.initial_mi() self.ini_j_all, self.ini_i_all = zip(*list(set(zip(self.ini_i, self.ini_j)))) if len(self.ini_j_all) == len(self.ini_i): print "Calculation based on small images may be leading to instablility. Choose a larger MAXIMUM or small DOWNSAMPLE." store, mistore = [], [] for self.ini_j, self.ini_i in zip(self.ini_j_all, self.ini_i_all): self.mi_last() store.append((self._j, self._i)) mistore.append(self.mi) j, i = store[mistore.index(max(mistore))] self.store = store self.mistore = mistore self._j, self._i, self.mi = j, i, max(mistore) if __name__ == "__main__": pass PK!:Sxcelltk/utils/miopt_align.pyfrom __future__ import print_function import SimpleITK as sitk from _mutinfo import mutual_information import numpy as np def offset_slice(pixels1, pixels2, i, j): '''Return two sliced arrays where the first slice is offset by i,j relative to the second slice. ''' if i < 0: height = min(pixels1.shape[0] + i, pixels2.shape[0]) p1_imin = -i p2_imin = 0 else: height = min(pixels1.shape[0], pixels2.shape[0] - i) p1_imin = 0 p2_imin = i p1_imax = p1_imin + height p2_imax = p2_imin + height if j < 0: width = min(pixels1.shape[1] + j, pixels2.shape[1]) p1_jmin = -j p2_jmin = 0 else: width = min(pixels1.shape[1], pixels2.shape[1] - j) p1_jmin = 0 p2_jmin = j p1_jmax = p1_jmin + width p2_jmax = p2_jmin + width p1 = pixels1[p1_imin:p1_imax, p1_jmin:p1_jmax] p2 = pixels2[p2_imin:p2_imax, p2_jmin:p2_jmax] return p1, p2 def optimize_mi(pixels1, pixels2, bins=256, i=0, j=0): p2, p1 = offset_slice(pixels2, pixels1, i, j) best = mutual_information(p1, p2, bins) while True: last_i = i last_j = j for new_i in range(last_i - 1, last_i + 2): for new_j in range(last_j - 1, last_j + 2): if new_i == 0 and new_j == 0: continue p2, p1 = offset_slice(pixels2, pixels1, new_i, new_j) if p1.any() and p2.any(): info = mutual_information(p1, p2, bins) else: info = 0 if info > best: best = info i = new_i j = new_j if i == last_i and j == last_j: return i, j, best def calc_crop_coordinates(store, shapes): """ Calculate how each images should be cropped. Take output from calc_jitters_multiple and return (start, end) coordinates for x, y for each image. """ max_w = max(i[1] for i in store) start_w = [max_w - i[1] for i in store] size_w = min([shapes[1] + i[1] for i in store]) - max_w max_h = max(i[0] for i in store) start_h = [max_h - i[0] for i in store] size_h = min([shapes[0] + i[0] for i in store]) - max_h return [(hi, hi+size_h, wi, wi+size_w) for hi, wi in zip(start_h, start_w)] def sitk_translation(img0, img1, off0=0, off1=0): """ + off1 ^ | off0 + <--- + ---> - off0 | v - off1 """ s0, s1 = sitk.GetImageFromArray(img0), sitk.GetImageFromArray(img1) s0, s1 = sitk.Cast(s0, sitk.sitkFloat32), sitk.Cast(s1, sitk.sitkFloat32) R = sitk.ImageRegistrationMethod() R.SetMetricAsMattesMutualInformation(numberOfHistogramBins=250) R.SetOptimizerAsRegularStepGradientDescent(4.0, .01, 200) cc = sitk.TranslationTransform(s0.GetDimension()) cc.SetOffset([-off1, -off0]) R.SetInitialTransform(cc) R.SetInterpolator(sitk.sitkLinear) R.SetShrinkFactorsPerLevel([4, 2, 1]) R.SetSmoothingSigmasPerLevel([8, 4, 1]) outTx = R.Execute(s0, s1) return -int(round(outTx.GetParameters()[1])), -int(round(outTx.GetParameters()[0])) def register_multiseeds(img0, img1, bins=250, initial=(-30, 0, 30)): store = [] for i in initial: for ii in initial: s0, s1 = sitk_translation(img0, img1, i, ii) p2, p1 = offset_slice(img1, img0, s0, s1) store.append(((s0, s1), mutual_information(p1, p2, bins))) store.sort(key=lambda x: x[1]) return store[-1] def _crop_sample_two(num, x, y, patch_h=200, patch_w=200): """ """ patch_h = patch_h if patch_h % 2 else patch_h + 1 patch_w = patch_w if patch_w % 2 else patch_w + 1 coords = ([np.random.randint(patch_h, x.shape[0] - patch_h) for i in range(num)], [np.random.randint(patch_w, x.shape[1] - patch_w) for i in range(num)]) h, w = int(np.floor(patch_h/2)), int(np.floor(patch_w/2)) xstack = np.zeros((num, patch_h, patch_w), np.float32) ystack = xstack.copy() for n, (ch, cw) in enumerate(zip(*coords)): xstack[n, :, :] = x[ch-h:ch+h+1, cw-w:cw+w+1] ystack[n, :, :] = y[ch-h:ch+h+1, cw-w:cw+w+1] return xstack, ystack def register_multiseeds_crop(img0, img1, bins=250, initial=(-30, 0, 30), num=5, patch_size=200): """ Align two images and return jitters and the highest mutual information it finds. It will crop an image into a (patch_size x patch_size) randomly and search for jitters until it finds the same values twice or it reaches num attempts. patch_size at least 150 is recommended. """ img0s, img1s = _crop_sample_two(num, img0, img1, patch_size, patch_size) st = [] for img0, img1 in zip(img0s, img1s): res = register_multiseeds(img0, img1, bins=bins, initial=initial) if [i for i in st if res[0] == i[0]]: return res st.append(res) print('No overlap. It may not be the optimal alignment') return sorted(st, key=lambda x:x[1])[-1] PK!r..celltk/utils/morphsnakes.py# -*- coding: utf-8 -*- """ morphsnakes =========== This is a Python implementation of the algorithms introduced in the paper Márquez-Neila, P., Baumela, L., Álvarez, L., "A morphological approach to curvature-based evolution of curves and surfaces". IEEE Transactions on Pattern Analysis and Machine Intelligence (PAMI), 2013. This implementation is intended to be as brief, understandable and self-contained as possible. It does not include any enhancement to make it fast or efficient. Any practical implementation of this algorithm should work only over the neighbor pixels of the 0.5-levelset, not over all the embedding function, and perhaps should feature multi-threading or GPU capabilities. The classes MorphGAC and MorphACWE provide most of the functionality of this module. They implement the Morphological Geodesic Active Contours and the Morphological Active Contours without Edges, respectively. See the aforementioned paper for full details. See test.py for examples of usage. """ __author__ = "P. Márquez Neila " from itertools import cycle import numpy as np from scipy import ndimage from scipy.ndimage import binary_dilation, binary_erosion, \ gaussian_filter, gaussian_gradient_magnitude class fcycle(object): def __init__(self, iterable): """Call functions from the iterable each time it is called.""" self.funcs = cycle(iterable) def __call__(self, *args, **kwargs): f = next(self.funcs) return f(*args, **kwargs) # SI and IS operators for 2D and 3D. _P2 = [np.eye(3), np.array([[0,1,0]]*3), np.flipud(np.eye(3)), np.rot90([[0,1,0]]*3)] _P3 = [np.zeros((3,3,3)) for i in range(9)] _P3[0][:,:,1] = 1 _P3[1][:,1,:] = 1 _P3[2][1,:,:] = 1 _P3[3][:,[0,1,2],[0,1,2]] = 1 _P3[4][:,[0,1,2],[2,1,0]] = 1 _P3[5][[0,1,2],:,[0,1,2]] = 1 _P3[6][[0,1,2],:,[2,1,0]] = 1 _P3[7][[0,1,2],[0,1,2],:] = 1 _P3[8][[0,1,2],[2,1,0],:] = 1 _aux = np.zeros((0)) def SI(u): """SI operator.""" global _aux if np.ndim(u) == 2: P = _P2 elif np.ndim(u) == 3: P = _P3 else: raise ValueError("u has an invalid number of dimensions (should be 2 or 3)") if u.shape != _aux.shape[1:]: _aux = np.zeros((len(P),) + u.shape) for i in range(len(P)): _aux[i] = binary_erosion(u, P[i]) return _aux.max(0) def IS(u): """IS operator.""" global _aux if np.ndim(u) == 2: P = _P2 elif np.ndim(u) == 3: P = _P3 else: raise ValueError("u has an invalid number of dimensions (should be 2 or 3)") if u.shape != _aux.shape[1:]: _aux = np.zeros((len(P),) + u.shape) for i in range(len(P)): _aux[i] = binary_dilation(u, P[i]) return _aux.min(0) # SIoIS operator. SIoIS = lambda u: SI(IS(u)) ISoSI = lambda u: IS(SI(u)) curvop = fcycle([SIoIS, ISoSI]) # Stopping factors (function g(I) in the paper). def gborders(img, alpha=1.0, sigma=1.0): """Stopping criterion for image borders.""" # The norm of the gradient. gradnorm = gaussian_gradient_magnitude(img, sigma, mode='constant') return 1.0/np.sqrt(1.0 + alpha*gradnorm) def glines(img, sigma=1.0): """Stopping criterion for image black lines.""" return gaussian_filter(img, sigma) class MorphACWE(object): """Morphological ACWE based on the Chan-Vese energy functional.""" def __init__(self, data, smoothing=1, lambda1=1, lambda2=1): """Create a Morphological ACWE solver. Parameters ---------- data : ndarray The image data. smoothing : scalar The number of repetitions of the smoothing step (the curv operator) in each iteration. In other terms, this is the strength of the smoothing. This is the parameter µ. lambda1, lambda2 : scalars Relative importance of the inside pixels (lambda1) against the outside pixels (lambda2). """ self._u = None self.smoothing = smoothing self.lambda1 = lambda1 self.lambda2 = lambda2 self.data = data def set_levelset(self, u): self._u = np.double(u) self._u[u>0] = 1 self._u[u<=0] = 0 levelset = property(lambda self: self._u, set_levelset, doc="The level set embedding function (u).") def step(self): """Perform a single step of the morphological Chan-Vese evolution.""" # Assign attributes to local variables for convenience. u = self._u if u is None: raise ValueError("the levelset function is not set (use set_levelset)") data = self.data # Determine c0 and c1. inside = u>0 outside = u<=0 c0 = data[outside].sum() / float(outside.sum()) c1 = data[inside].sum() / float(inside.sum()) # Image attachment. dres = np.array(np.gradient(u)) abs_dres = np.abs(dres).sum(0) #aux = abs_dres * (c0 - c1) * (c0 + c1 - 2*data) aux = abs_dres * (self.lambda1*(data - c1)**2 - self.lambda2*(data - c0)**2) res = np.copy(u) res[aux < 0] = 1 res[aux > 0] = 0 # Smoothing. for i in range(self.smoothing): res = curvop(res) self._u = res def run(self, iterations): """Run several iterations of the morphological Chan-Vese method.""" for i in range(iterations): self.step() class MorphGAC(object): """Morphological GAC based on the Geodesic Active Contours.""" def __init__(self, data, smoothing=1, threshold=0, balloon=0): """Create a Morphological GAC solver. Parameters ---------- data : array-like The stopping criterion g(I). See functions gborders and glines. smoothing : scalar The number of repetitions of the smoothing step in each iteration. This is the parameter µ. threshold : scalar The threshold that determines which areas are affected by the morphological balloon. This is the parameter θ. balloon : scalar The strength of the morphological balloon. This is the parameter ν. """ self._u = None self._v = balloon self._theta = threshold self.smoothing = smoothing self.set_data(data) def set_levelset(self, u): self._u = np.double(u) self._u[u>0] = 1 self._u[u<=0] = 0 def set_balloon(self, v): self._v = v self._update_mask() def set_threshold(self, theta): self._theta = theta self._update_mask() def set_data(self, data): self._data = data self._ddata = np.gradient(data) self._update_mask() # The structure element for binary dilation and erosion. self.structure = np.ones((3,)*np.ndim(data)) def _update_mask(self): """Pre-compute masks for speed.""" self._threshold_mask = self._data > self._theta self._threshold_mask_v = self._data > self._theta/np.abs(self._v) levelset = property(lambda self: self._u, set_levelset, doc="The level set embedding function (u).") data = property(lambda self: self._data, set_data, doc="The data that controls the snake evolution (the image or g(I)).") balloon = property(lambda self: self._v, set_balloon, doc="The morphological balloon parameter (ν (nu, not v)).") threshold = property(lambda self: self._theta, set_threshold, doc="The threshold value (θ).") def step(self): """Perform a single step of the morphological snake evolution.""" # Assign attributes to local variables for convenience. u = self._u gI = self._data dgI = self._ddata theta = self._theta v = self._v if u is None: raise ValueError("the levelset is not set (use set_levelset)") res = np.copy(u) # Balloon. if v > 0: aux = binary_dilation(u, self.structure) elif v < 0: aux = binary_erosion(u, self.structure) if v!= 0: res[self._threshold_mask_v] = aux[self._threshold_mask_v] # Image attachment. aux = np.zeros_like(res) dres = np.gradient(res) for el1, el2 in zip(dgI, dres): aux += el1*el2 res[aux > 0] = 1 res[aux < 0] = 0 # Smoothing. for i in range(self.smoothing): res = curvop(res) self._u = res def run(self, iterations): """Run several iterations of the morphological snakes method.""" for i in range(iterations): self.step() def evolve_visual(msnake, levelset=None, num_iters=20, background=None): """ Visual evolution of a morphological snake. Parameters ---------- msnake : MorphGAC or MorphACWE instance The morphological snake solver. levelset : array-like, optional If given, the levelset of the solver is initialized to this. If not given, the evolution will use the levelset already set in msnake. num_iters : int, optional The number of iterations. background : array-like, optional If given, background will be shown behind the contours instead of msnake.data. """ from matplotlib import pyplot as ppl if levelset is not None: msnake.levelset = levelset # Prepare the visual environment. fig = ppl.gcf() fig.clf() ax1 = fig.add_subplot(1,2,1) if background is None: ax1.imshow(msnake.data, cmap=ppl.cm.gray) else: ax1.imshow(background, cmap=ppl.cm.gray) ax1.contour(msnake.levelset, [0.5], colors='r') ax2 = fig.add_subplot(1,2,2) ax_u = ax2.imshow(msnake.levelset) ppl.pause(0.001) # Iterate. for i in range(num_iters): # Evolve. msnake.step() # Update figure. del ax1.collections[0] ax1.contour(msnake.levelset, [0.5], colors='r') ax_u.set_data(msnake.levelset) fig.canvas.draw() #ppl.pause(0.001) # Return the last levelset. return msnake.levelset def evolve_visual3d(msnake, levelset=None, num_iters=20): """ Visual evolution of a three-dimensional morphological snake. Parameters ---------- msnake : MorphGAC or MorphACWE instance The morphological snake solver. levelset : array-like, optional If given, the levelset of the solver is initialized to this. If not given, the evolution will use the levelset already set in msnake. num_iters : int, optional The number of iterations. """ from mayavi import mlab import matplotlib.pyplot as ppl if levelset is not None: msnake.levelset = levelset fig = mlab.gcf() mlab.clf() src = mlab.pipeline.scalar_field(msnake.data) mlab.pipeline.image_plane_widget(src, plane_orientation='x_axes', colormap='gray') cnt = mlab.contour3d(msnake.levelset, contours=[0.5]) @mlab.animate(ui=True) def anim(): for i in range(num_iters): msnake.step() cnt.mlab_source.scalars = msnake.levelset print("Iteration %s/%s..." % (i + 1, num_iters)) yield anim() mlab.show() # Return the last levelset. return msnake.levelsetPK!y~celltk/utils/pairwise.pyimport numpy as np def one_to_one_assignment(binary_cost, value_cost): '''When mulitple True exists in row or column, it will pick one with the lowest value_cost. ''' binary_cost = pick_closer(binary_cost, value_cost) binary_cost = pick_closer(binary_cost.T, value_cost.T) binary_cost = binary_cost.T return binary_cost def one_to_two_assignment(binary_cost, value_cost): '''If there are more than two True in a row, make them to two. ''' # First make sure the daughter is not shared by two parents binary_cost = pick_closer(binary_cost.T, value_cost.T) binary_cost = binary_cost.T # pick two based on value_cost binary_cost = pick_closer_two(binary_cost, value_cost) return binary_cost def find_one_to_one_assign(binary_cost): cost = binary_cost.copy() (_, col1) = np.where([np.sum(cost, 0) == 1]) cost[np.sum(cost, 1) != 1] = False (row, col2) = np.where(cost) good_row = [ci for ri, ci in zip(row, col2) if ci in col1] good_col = [ri for ri, ci in zip(row, col2) if ci in good_row] return good_row, good_col def pick_closer(binary_cost, value_cost): '''If there are several True in a row of binary_cost, it will pick one with the lowest value_cost. If mulitple elements have the same value_cost, it will pick the first one. ''' for x in range(binary_cost.shape[0]): binary_row = binary_cost[x, :] value_row = value_cost[x, :] if binary_row.any(): min_value = np.min(value_row[binary_row]) idx = np.where(value_row == min_value)[0][0] binary_row[0:idx] = False binary_row[idx+1:] = False return binary_cost def pick_closer_two(binary_cost, value_cost, PICK=2): for x in range(binary_cost.shape[0]): binary_row = binary_cost[x, :] value_row = value_cost[x, :] if binary_row.sum() > 1: binary_row_copy = binary_row.copy() sorted_idx = np.argsort(value_row[binary_row]) binary_row[:] = False for i in sorted_idx[:PICK]: idx = np.where(value_row == value_row[binary_row_copy][i])[0][0] binary_row[idx] = True return binary_cost PK!|OBcelltk/utils/parser.pyimport re import ast import __builtin__ import ast import argparse def split_params(inputs): if "/" not in inputs: return [inputs] store = [] li = [] while inputs: element = inputs.pop(0) if element == "/": store.append(li) li = [] else: li.append(element) store.append(li) return store class ParamParser(object): def __init__(self, param_args): self.param_args = param_args def run(self): if self.param_args is None: return [{}] parameters = split_params(self.param_args[0]) return [self.convert2dict(p) for p in parameters] def convert2dict(self, param): dictargs = dict(e.split('=') for e in param) for key, value in dictargs.iteritems(): if value[0].isdigit(): dictargs[key] = ast.literal_eval(value) elif value[0]=='[': try: dictargs[key] = ast.literal_eval(value) except: temp = [] for i in value[1:-1].split(','): if i.isdigit(): temp.append(ast.literal_eval(i)) else: temp.append(i) dictargs[key] = temp return dictargs class ParamParser1(object): def __init__(self, param_args): self.param_args = param_args def run(self): params = self.split_params(self.param_args[:]) params = self.iter_combine_list(params) param_list = [] for param in params: param_list.append(self.add_quotation(param)) dict_list = [dict(i) for i in param_list] return self.iter_literal(dict_list) def iter_literal(self, dict_list): for dic in dict_list: for key, value in dic.iteritems(): dic[key] = ast.literal_eval(value) return dict_list def iter_combine_list(self, params): store = [] for param in params: store.append(self.combine_list(param)) return store def add_quotation(self, ps): """ """ pss = [] for p in ps: pair = p.split("=") if not pair[1] in dir(__builtin__): pair[1] = re.sub('([a-zA-Z/_][a-zA-Z0-9/._\*]*)', "'\g<1>'", pair[1]) pss.append(pair) return pss def split_params(self, inputs): if "/" not in inputs: return [inputs] store = [] li = [] while inputs: element = inputs.pop(0) if element == "/": store.append(li) li = [] else: li.append(element) store.append(li) return store def combine_list(self, param): if len(param) == 1: return param itr = iter(param) ps = [] while True: try: p = itr.next() if "[" in p: temp = [] temp.append(p) while True: p = itr.next() temp.append(p) if "]" in p: break ps.append("".join(temp)) else: ps.append(p) except StopIteration: break return ps def parse_image_files(inputs): if "/" not in inputs: return inputs store = [] li = [] while inputs: element = inputs.pop(0) if element == "/": store.append(li) li = [] else: li.append(element) store.append(li) return zip(*store)PK!ߐVV!celltk/utils/postprocess_utils.pyfrom skimage.measure._regionprops import _RegionProperties import numpy as np import scipy.ndimage as ndi from global_holder import holder from math import sqrt from filters import label class _RegionProperties2(_RegionProperties): parent = None nxt = None def __init__(self, slice, label, label_image, intensity_image, cache_active): if intensity_image is not None: if not intensity_image.shape == label_image.shape: raise ValueError('Label and intensity image must have the' 'same shape.') self.label = label self._slice = slice self.slice = slice self._label_image = label_image self._intensity_image = intensity_image self._cache_active = cache_active self._cache = {} self._ndim = label_image.ndim # Note: in PR 2603, we added support for nD moments in regionprops. # Many properties used xy coordinates, instead of rc. This attribute # helps with the deprecation process and should be removed in 0.16. self._use_xy_warning = True self._transpose_moments = True @property def total_intensity(self): return np.sum(self.intensity_image[self.image]).astype(np.float) @property def x(self): return self.centroid[1] @property def y(self): return self.centroid[0] @property def median_intensity(self): return np.median(self.intensity_image[self.image]).astype(np.float) @property def std_intensity(self): return np.std(self.intensity_image[self.image]).astype(np.float) @property def cv_intensity(self): return (self.std_intensity/self.mean_intensity).astype(np.float) @property def cell_id(self): return self.label @property def minor_axis_length(self): _, l2 = self.inertia_tensor_eigvals return 4 * sqrt(max(l2, 0)) @property def major_axis_length(self): l1, _ = self.inertia_tensor_eigvals return 4 * sqrt(max(l1, 0)) @property def num_seg(self): """ number of isolated segments """ return np.max(label(self.image, connectivity=1)) def regionprops(label_image, intensity_image=None, cache=True): label_image = np.squeeze(label_image) if label_image.ndim not in (2, 3): raise TypeError('Only 2-D and 3-D images supported.') if not np.issubdtype(label_image.dtype, np.integer): raise TypeError('Label image must be of integral type.') regions = [] objects = ndi.find_objects(label_image) for i, sl in enumerate(objects): if sl is None: continue label = i + 1 props = _RegionProperties2(sl, label, label_image, intensity_image, cache) regions.append(props) return regions class Cell(object): def __init__(self, regionprop): for p in dir(regionprop): if "__" in p: continue ret = getattr(regionprop, p) if not isinstance(ret, dict) and not isinstance(ret, tuple) and not isinstance(ret, np.ndarray): setattr(self, p, ret) self.coords = regionprop.coords self.abs_id = holder.count() self.centroid = regionprop.centroid class LCell(object): def __init__(self, regionprop): for p in ['area', 'num_seg', 'median_intensity', 'total_intensity', 'x', 'y', 'cell_id', 'label']: setattr(self, p, getattr(regionprop, p)) self.abs_id = holder.count() self._original_label = self.label self.nxt = None self.parent = None # class LCell(object): # def __init__(self, regionprop): # self._labels = ['area', 'num_seg', 'median_intensity', 'total_intensity', 'x', 'y', 'cell_id', 'label'] # self._data = np.zeros(len(self._labels)) # for num, p in enumerate(self._labels): # self._data[num] = getattr(regionprop, p) # self.parent = None # self.nxt =None # def __getattr__(self, name): # if name in self._labels: # return self._data[self._labels.index(name)] # super(LCell, self).__getattr__(name) # def __setattr__(self, name, value): # object.__setattr__(self, name, value) # if name in self._labels: # self._data[self._labels.index(name)] = value PK!x celltk/utils/preprocess_utils.pyimport numpy as np from skimage.transform import resize from pywt import WaveletPacket2D import SimpleITK as sitk from wavelet_bgr import WaveletBGR from rolling_ball import PyRollingBall from functools import partial from skimage.transform import resize from scipy.ndimage.filters import gaussian_filter def adaptive_thresh(img, RATIO=3.0, FILTERINGSIZE=50): """Segment as a foreground if pixel is higher than ratio * blurred image. If you set ratio 3.0, it will pick the pixels 300 percent brighter than the blurred image. """ fim = gaussian_filter(img, FILTERINGSIZE) bw = img > (fim * RATIO) return bw def histogram_matching(img, previmg, BINS=500, QUANT=2, THRES=False): simg = sitk.GetImageFromArray(img) spimg = sitk.GetImageFromArray(previmg) fil = sitk.HistogramMatchingImageFilter() fil.SetNumberOfHistogramLevels(BINS) fil.SetNumberOfMatchPoints(QUANT) fil.SetThresholdAtMeanIntensity(THRES) filimg = fil.Execute(simg, spimg) return sitk.GetArrayFromImage(filimg) def estimate_background_prc(img, BLOCK, PERCENTILE): im = img.copy() im = im[int(im.shape[0]*0.1):int(im.shape[0]*0.9), int(im.shape[1]*0.1):int(im.shape[1]*0.9)] xSize, ySize = im.shape x = np.linspace(0, xSize, BLOCK+1, dtype=int) y = np.linspace(0, ySize, BLOCK+1, dtype=int) tempim = np.zeros((BLOCK, BLOCK)) for xn, (xs, xe) in enumerate(zip(x[:-1], x[1:])): for yn, (ys, ye) in enumerate(zip(y[:-1], y[1:])): blkim = im[xs:xe, ys:ye] tempim[xn, yn] = np.percentile(blkim, PERCENTILE) background = resize(tempim, img.shape, order=3, mode='reflect') img = img - background return img def convert_positive(img, OFFSET=50): """OFFSET prevents the error when transforming image to log scale. """ img[img < OFFSET] = OFFSET # img = img + OFFSET return img def wavelet_subtraction(img, level): """6- 7 level is recommended""" if level == 0: return img wp = WaveletPacket2D(data=img, wavelet='haar', mode='sym') back = resize(np.array(wp['a'*level].data), img.shape, order=3, mode='reflect')/(2**level) img = img - back return img def homogenize_intensity_n4(img, background_bw): simg = sitk.GetImageFromArray(img.astype(np.float32)) sbw = sitk.GetImageFromArray((background_bw).astype(np.uint8)) fil = sitk.N4BiasFieldCorrectionImageFilter() cimg = fil.Execute(simg, sbw) return sitk.GetArrayFromImage(cimg) def curvature_anisotropic_smooth(img, NUMITER=10): fil = sitk.CurvatureAnisotropicDiffusionImageFilter() fil.SetNumberOfIterations(NUMITER) simg = sitk.GetImageFromArray(img.astype(np.float32)) sres = fil.Execute(simg) return sitk.GetArrayFromImage(sres) def remove_odd_addback(img, func): rshape, cshape = img.shape if not rshape % 2 == 0: img = img[:-1, :] if not cshape % 2 == 0: img = img[:, :-1] img = func(img) if not rshape % 2 == 0: output = np.zeros((img.shape[0]+1, img.shape[1])) output[:-1, :] = img output[-1, :] = img[-1, :] img = output if not cshape % 2 == 0: output = np.zeros((img.shape[0], img.shape[1]+1)) output[:, :-1] = img output[:, -1] = img[:, -1] img = output return img def wavelet_subtraction_hazen(img, ITER=5, THRES=100, WLEVEL=6): wb = WaveletBGR(padding_mode='sym') f = partial(wb.estimateBG, iterations=ITER, threshold=THRES, wavelet_level=WLEVEL) background = remove_odd_addback(img, f) return background def resize_img(himg, origshape): """resize works for float range from 0 to 1. """ himg = np.float32(himg) minh = himg.min() cimg = himg - minh maxh = cimg.max() cimg = cimg/maxh resized = resize(cimg, origshape) resized *= maxh resized += minh return resized def rolling_ball_subtraction_hazen(img, RADIUS=10, SIGMA=3): rb = PyRollingBall(ball_radius=RADIUS, smoothing_sigma=SIGMA) f = partial(rb.estimateBG) background = remove_odd_addback(img, f) return background PK!{rf* * celltk/utils/rolling_ball.pyimport numpy import scipy import scipy.ndimage # from numba import double from numba.decorators import jit @jit def estimatejit(image, sm_image, ball_image, ball, ball_size, ball_radius): for x in range(image.shape[0]): for y in range(image.shape[1]): min_z = sm_image[x,y] for bx in range(2*ball_size+1): cx = x + bx - ball_size if (cx < 0): continue if (cx >= image.shape[0]): continue for by in range(2*ball_size+1): cy = y + by - ball_size if (cy < 0): continue if (cy >= image.shape[1]): continue cz = sm_image[cx,cy] - ball[bx,by] if (cz < min_z): min_z = cz ball_image[x,y] = min_z ball_image += ball_radius return ball_image class PyRollingBall(object): """ Rolling ball smoothing class. """ def __init__(self, ball_radius, smoothing_sigma): """ ball_radius is the ball radius in pixels. smoothing_sigma is the sigma of the gaussian to use for image smoothing prior estimating the background using the rolling ball. This assumes that the ball is relatively large compared to variations in the surface topology. """ self.ball_radius = ball_radius self.smoothing_sigma = smoothing_sigma self.ball_size = int(round(ball_radius * 0.5)) br = ball_radius * ball_radius self.ball = numpy.zeros((2*self.ball_size+1, 2*self.ball_size+1)) for x in range(2*self.ball_size+1): dx = x - self.ball_size for y in range(2*self.ball_size+1): dy = y - self.ball_size self.ball[x,y] = br - (dx * dx + dy * dy) self.ball = numpy.sqrt(self.ball) def estimateBG(self, image): image = image.astype(numpy.float) sm_image = scipy.ndimage.filters.gaussian_filter(image, self.smoothing_sigma) ball_image = numpy.zeros(image.shape) return estimatejit(image, sm_image, ball_image, self.ball, self.ball_size, self.ball_radius) def removeBG(self, image): return image - self.estimateBG(image) PK!6   "celltk/utils/shading_correction.pyfrom __future__ import division import numpy as np import json import tempfile import urllib import shutil from os.path import join import os from util import imread #from utils.util import imread from glob import glob import tifffile as tiff def retrieve_ff_ref(refpath, darkrefpath): """ refpath: 'http://archive.simtk.org/ktrprotocol/temp/ffref_20x3bin.npz' darkrefpath: 'http://archive.simtk.org/ktrprotocol/temp/ffdarkref_20x3bin.npz' """ try: temp_dir = tempfile.mkdtemp() urllib.urlretrieve(refpath, join(temp_dir, 'ref.npz')) ref = np.load(join(temp_dir, 'ref.npz')) urllib.urlretrieve(darkrefpath, join(temp_dir, 'darkref.npz')) darkref = np.load(join(temp_dir, 'darkref.npz')) finally: shutil.rmtree(temp_dir) # delete directory return ref, darkref def correct_shade(img, ref, darkref, ch): img = img.astype(np.float) d0 = img.astype(np.float) - darkref[ch] d1 = ref[ch] - darkref[ch] return d1.mean() * d0/d1 def shading_correction_folder(inputfolder, outputfolder, binning=3, magnification=20): """ Covert lab specific. Not to be called by preprocess_operation. Use it in separate from a pipeline. """ refpath = 'http://archive.simtk.org/ktrprotocol/temp/ffref_{0}x{1}bin.npz'.format(magnification, binning) darkrefpath = 'http://archive.simtk.org/ktrprotocol/temp/ffdarkref_{0}x{1}bin.npz'.format(magnification, binning) ref, darkref = retrieve_ff_ref(refpath, darkrefpath) parentfolder = inputfolder for dirname, subdirlist, filelist in os.walk(parentfolder): if 'metadata.txt' in filelist: sfol_name = dirname.split(parentfolder)[-1] sfol_name = sfol_name if not sfol_name.startswith('/') else sfol_name[1:] outputdir = join(outputfolder, sfol_name) if not os.path.exists(outputdir): os.makedirs(outputdir) with open(join(dirname, 'metadata.txt')) as mfile: data = json.load(mfile) channels = data['Summary']['ChNames'] for chnum, ch in enumerate(channels): pathlist = glob(join(dirname, '*channel{0:03d}*'.format(chnum))) for path in pathlist: try: img = correct_shade(imread(path), ref, darkref, ch) except: print "ch might not exist as a reference." img = imread(path) tiff.imsave(join(outputdir, os.path.basename(path)), img.astype(np.float32)) PK!PPcelltk/utils/stitch_utils.pyimport numpy as np """ Stitch images with Fiji results Exported from Stitch_image_Grid_Sequence in Fiji e.g. points = [(0.0, 0.0), (-556.46246, -8.537842), (-565.82874, 533.8285), (-131.09012, 483.0899)] """ def relative_position(points): points_flat = [int(el) for inner in points for el in inner] xpoints = points_flat[1::2] ypoints = points_flat[0::2] max_xp = min(xpoints) max_yp = min(ypoints) rel_xpoints = map(lambda l: l - max_xp, xpoints) rel_ypoints = map(lambda l: l - max_yp, ypoints) rel_points = (rel_xpoints, rel_ypoints) return rel_points def replace_peri(img, val=0): img[0, :] = val img[-1, :] = val img[:, 0] = val img[:, -1] = val return img def stitching(img, rel_points): imShape=img.shape rel_xpoints=rel_points[0] rel_ypoints=rel_points[1] stitchImg = np.zeros((max(rel_xpoints)+imShape[0], max(rel_ypoints)+imShape[1], imShape[2])) for pos in range(imShape[2]): tmp = img[:, :, pos] stitchImg[rel_xpoints[pos]:rel_xpoints[pos]+imShape[0], rel_ypoints[pos]:rel_ypoints[pos]+imShape[1], pos] = tmp stitchImg[:,:,-1] = np.array(stitchImg.max(axis=2), dtype=np.float32) stitchImg = stitchImg[sorted(rel_xpoints)[1]:imShape[0]+sorted(rel_xpoints)[2], sorted(rel_ypoints)[1]:imShape[0]+sorted(rel_ypoints)[2]] return stitchImg PK!bbcelltk/utils/subdetect_utils.pyimport SimpleITK as sitk import numpy as np from scipy.ndimage import distance_transform_edt from skimage.morphology import watershed as skiwatershed from skimage.measure import label from skimage.feature import peak_local_max from skimage.morphology import disk from skimage.morphology import closing from skimage.measure import label as skim_label from scipy.ndimage.filters import gaussian_filter from scipy.signal import convolve2d from skimage.measure import regionprops def dilate_sitk(labels, RAD): slabels = sitk.GetImageFromArray(labels) gd = sitk.GrayscaleDilateImageFilter() gd.SetKernelRadius(RAD) return sitk.GetArrayFromImage(gd.Execute(slabels)) def voronoi_expand(labels, return_line=False): dist = distance_transform_edt(labels) vor = skiwatershed(-dist, markers=labels) if not return_line: return vor else: mask = skiwatershed(-dist, markers=labels, watershed_line=True) lines = mask == 0 return vor, lines def calc_mask_exclude_overlap(nuclabel, RINGWIDTH=5): """ Examples: >>> template = np.zeros((5, 5)) >>> template[1, 1] = 1 >>> template[-2, -2] = 2 >>> calc_mask_exclude_overlap(template, 2) array([[False, False, False, False, False], [False, False, True, True, False], [False, True, True, True, False], [False, True, True, False, False], [False, False, False, False, False]], dtype=bool) """ dilated_nuc = dilate_sitk(nuclabel.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 6e4 - nuclabel comp_dilated_nuc[comp_dilated_nuc == 6e4] = 0 comp_dilated_nuc = dilate_sitk(comp_dilated_nuc.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 6e4 - comp_dilated_nuc comp_dilated_nuc[comp_dilated_nuc == 6e4] = 0 mask = comp_dilated_nuc != dilated_nuc return mask def dilate_to_cytoring(labels, RINGWIDTH, MARGIN): """ Examples: >>> template = np.zeros((5, 5)) >>> template[2, 2] = 1 >>> dilate_to_cytoring(template, 1, 0) array([[0, 0, 0, 0, 0], [0, 1, 1, 1, 0], [0, 1, 0, 1, 0], [0, 1, 1, 1, 0], [0, 0, 0, 0, 0]], dtype=uint16) >>> dilate_to_cytoring(template, 2, 1) array([[0, 1, 1, 1, 0], [1, 0, 0, 0, 1], [1, 0, 0, 0, 1], [1, 0, 0, 0, 1], [0, 1, 1, 1, 0]], dtype=uint16) """ dilated_nuc = dilate_sitk(labels.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 1e4 - labels comp_dilated_nuc[comp_dilated_nuc == 1e4] = 0 comp_dilated_nuc = dilate_sitk(comp_dilated_nuc.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 1e4 - comp_dilated_nuc comp_dilated_nuc[comp_dilated_nuc == 1e4] = 0 dilated_nuc[comp_dilated_nuc != dilated_nuc] = 0 if MARGIN == 0: antinucmask = labels else: antinucmask = dilate_sitk(np.int32(labels), MARGIN) dilated_nuc[antinucmask.astype(bool)] = 0 return dilated_nuc.astype(np.uint16) def dilate_to_cytoring_buffer(labels, RINGWIDTH, MARGIN, BUFFER): """ Examples: >>> template = np.zeros((5, 5)) >>> template[1, 1] = 1 >>> template[-2, -2] = 2 >>> dilate_to_cytoring_buffer(template, 2, 0, 1) array([[1, 1, 0, 0, 0], [1, 0, 0, 0, 0], [0, 0, 0, 0, 0], [0, 0, 0, 0, 2], [0, 0, 0, 2, 2]], dtype=uint16) """ dilated_nuc = dilate_sitk(labels.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 1e4 - labels comp_dilated_nuc[comp_dilated_nuc == 1e4] = 0 comp_dilated_nuc = dilate_sitk(comp_dilated_nuc.astype(np.int32), RINGWIDTH) comp_dilated_nuc = 1e4 - comp_dilated_nuc comp_dilated_nuc[comp_dilated_nuc == 1e4] = 0 vor, vlines = voronoi_expand(labels, return_line=True) vor0, vor1 = vor.copy(), vor.copy() vor0[comp_dilated_nuc != vor0.copy()] = False vor1[dilated_nuc != vor1.copy()] = False dilated_nuc = np.max(np.dstack((vor0, vor1)), axis=2) if MARGIN == 0: antinucmask = labels else: antinucmask = dilate_sitk(np.int32(labels), MARGIN) dilated_nuc[antinucmask.astype(bool)] = 0 if BUFFER: vlines = dilate_sitk(vlines.astype(np.uint32), BUFFER) dilated_nuc[vlines > 0] = 0 return dilated_nuc.astype(np.uint16) def watershed_labels(labels, regmax): # Since there are non-unique values for dist, add very small numbers. This will separate each marker by regmax at least. dist = distance_transform_edt(labels) + np.random.rand(*labels.shape)*1e-10 labeled_maxima = label(peak_local_max(dist, min_distance=int(regmax), indices=False)) wshed = -dist wshed = wshed - np.min(dist) markers = np.zeros(wshed.shape, np.int16) markers[labeled_maxima > 0] = -labeled_maxima[labeled_maxima > 0] markers[labels == 0] = 0 wlabels = skiwatershed(wshed, markers, connectivity=np.ones((3,3), bool), mask=labels!=0) wlabels = -wlabels wlabels = labels.max() + wlabels wlabels[wlabels == labels.max()] = 0 all_labels = label(labels + wlabels) return all_labels def pairwise_distance(loc1, loc2): xprev = [i[0] for i in loc1] yprev = [i[1] for i in loc1] xcurr = [i[0] for i in loc2] ycurr = [i[1] for i in loc2] xprevTile = np.tile(xprev, (len(xcurr), 1)) yprevTile = np.tile(yprev, (len(ycurr), 1)) return abs(xprevTile.T - xcurr) + abs(yprevTile.T - ycurr) def skilabel(bw, conn=2): '''original label might label any objects at top left as 1. To get around this pad it first.''' bw = np.pad(bw, pad_width=1, mode='constant', constant_values=False) label = skim_label(bw, connectivity=conn) label = label[1:-1, 1:-1] return label def calc_high_pass_kernel(slen, SIGMA): """For Salmonella""" temp = np.zeros((slen, slen)) temp[int(slen/2), int(slen/2)] = 1 gf = gaussian_filter(temp, SIGMA) norm = np.ones((slen, slen))/(slen**2) return gf - norm def calc_high_pass(img, slen, SIGMA): """For Salmonella""" kernel = calc_high_pass_kernel(slen, SIGMA) cc = convolve2d(img, kernel, mode='same') return cc def label_high_pass(img, slen=3, SIGMA=0.5, THRES=50, CLOSE=3): """For Salmonella""" cc = calc_high_pass(img, slen, SIGMA) cc[cc < 0] = 0 la = skilabel(cc > THRES, conn=1) la = closing(la, disk(CLOSE)) return la def label_nearest(img, label, nuc, DISTHRES=25): """Label objects to the nearest nuc. """ nuc_prop = regionprops(nuc, img, cache=False) sal_prop = regionprops(label, img, cache=False) nuc_loc = [i.centroid for i in regionprops(nuc, img, cache=False)] sal_loc = [i.centroid for i in regionprops(label, img, cache=False)] dist = pairwise_distance(nuc_loc, sal_loc) min_dist_arg = np.argmin(dist, axis=0) template = np.zeros(img.shape, np.uint16) for num, (idx, sal) in enumerate(zip(min_dist_arg, sal_prop)): if dist[idx, num] < DISTHRES: template[sal.coords[:, 0], sal.coords[:, 1]] = nuc_prop[idx].label comb = np.max(np.dstack((template, nuc)), axis=2).astype(np.uint16) return template, comb, nuc_prop, nuc_loc PK!y celltk/utils/tfutils.pyimport os import imp import numpy as np from scipy.ndimage import imread as imread0 import tifffile as tiff def conv_labels2dto3d(labels): lbnums = np.unique(labels) arr = np.zeros((labels.shape[0], labels.shape[1], len(lbnums)), np.uint8) for i in lbnums: arr[:, :, i] = labels == i return arr def normalize(orig_img): percentile = 99.9 high = np.percentile(orig_img, percentile) low = np.percentile(orig_img, 100-percentile) img = np.minimum(high, orig_img) img = np.maximum(low, img) img = (img - low) / (high - low) return img def make_outputdir(output): try: os.makedirs(output) except: pass def imread_check_tiff(path): img = imread0(path) if img.dtype == 'object' or path.endswith('tif'): img = tiff.imread(path) return img def imread(path): if isinstance(path, tuple) or isinstance(path, list): st = [] for p in path: st.append(imread_check_tiff(p)) img = np.dstack(st) if img.shape[2] == 1: np.squeeze(img, axis=2) return img else: return imread_check_tiff(path) def parse_image_files(inputs): if "/" not in inputs: return (inputs, ) store = [] li = [] while inputs: element = inputs.pop(0) if element == "/": store.append(li) li = [] else: li.append(element) store.append(li) return zip(*store) def pad_image(image): # assumes the image is an np array of dimensions d0 x d1 x d2 x d3 # where d1 is height, d2 is width, and d3 is colors # returns a list of padded image, hpadding (tuple), wpadding (tuple) height = image.shape[1] hdelta = 8 - (height % 8) if (hdelta == 8): hpadding = (0, 0) elif (hdelta % 2) == 0: hpadding = (int(hdelta/2.0), int(hdelta/2.0)) else: hpadding = (int(hdelta/2.0), int(hdelta/2.0)+1) width = image.shape[2] wdelta = 8 - (width % 8) if (wdelta == 8): wpadding = (0, 0) elif (wdelta % 2) == 0: wpadding = (int(wdelta/2.0), int(wdelta/2.0)) else: wpadding = (int(wdelta/2.0), int(wdelta/2.0)+1) return [np.pad(image, ((0, 0), hpadding, wpadding, (0, 0)), 'constant', constant_values=0.0), hpadding, wpadding] def normalize_predictions(predictions): # predictions is typically a 3 x h x w list predictions = np.array(predictions) num_rows = predictions.shape[1] num_cols = predictions.shape[2] for i in range(num_rows): for j in range(num_cols): prediction = predictions[:, i, j] if (prediction.sum() != 0): predictions[:, i, j] = prediction / prediction.sum() return predictions PK!4celltk/utils/traces.pyimport numpy as np from operator import attrgetter import copy from itertools import izip import sys from track_utils import calc_massdiff, calc_diff from scipy.spatial.distance import cdist from collections import deque sys.setrecursionlimit(10000) class TracesController(object): def __init__(self, traces): self.traces = traces frames = [cell.frame for cell in [i[-1] for i in traces]] self.last_frame = max(frames) def disappeared(self): cells = [i[-1] for i in self.traces] return [cell for cell in cells if cell.frame < self.last_frame] def appeared(self): cells = [i[0] for i in self.traces] return [cell for cell in cells if cell.frame > 0] def pairwise_dist(self): if self.disappeared() and self.appeared(): dist = cdist([(i.y, i.x) for i in self.disappeared()], [(i.y, i.x) for i in self.appeared()]) return dist else: return np.array([]) def pairwise_frame(self): '''disappeared in row, appeared in col''' return calc_diff([i.frame for i in self.disappeared()], [i.frame for i in self.appeared()]) def pairwise_mass(self): return calc_massdiff(self.disappeared(), self.appeared()) def label_traces(traces): idx = 1 for trace in traces: for cell in trace: cell.label = idx idx += 1 return traces def assign_next_and_abs_id_to_storage(storage): '''Set next if two cells in consecutive frames have the same label_id''' for prev_cells, curr_cells in zip(storage[0:-1], storage[1:]): for prev_cell in prev_cells: for curr_cell in curr_cells: if prev_cell.label == curr_cell.label: prev_cell.nxt = curr_cell storage = [i for j in storage for i in j] # set abs_id from 1 to max [setattr(si, 'abs_id', i+1) for i, si in enumerate(storage)] return storage def construct_traces_based_on_next(storage): ''' Convert storage to traces. traces is a list of lists containing cells with same label_id. If cell1.nxt = cell2 and cell2.nxt = cell3, cell4.nxt = cell5, then traces = [[cell1, cell2, cell3], [cell4, cell5]]. storage has to be sorted based on frame. ''' traces = [] for cell in storage: cells = [cell] while cell.nxt is not None: cell = cell.nxt cells.append(storage.pop(storage.index(cell))) traces.append(cells) return traces def construct_traces_based_on_prev(storage): ''' ''' for cell in storage: cell.nxt.prev = cell.label for cell in storage: cell.prev = cell.parent for cell in reversed(storage): cells = deque([cell]) while cell.prev is not None: cell = cell.prev cells.append(storage.pop(storage.index(cell))) traces.append(cells) return traces def convert_traces_to_storage(traces): '''Convert traces to storage. ''' storage = [i for j in traces for i in j] return sorted(storage, key=attrgetter('frame')) def connect_parent_daughters(traces): '''Make sure to use this after saving labels. Parents will be duplicated, and Cell_id will be updated Assume traces = [[cell2, cell3], [cell4, cell5]], [cell6, cell7]] and cell4.parent = cell3, cell6.parent = cell3. Then this converts traces to [[cell2, cell3, cell4, cell5], [cell2, cell3, cell6, cell7]]. ''' # traces = construct_traces_based_on_next(storage) # Concatenate parent and daughters parent_cells = [i[0].parent for i in traces if i[0].parent is not None] traces_without_parent = [] parental_traces = [] while traces: trace = traces.pop(0) if trace[-1] in parent_cells: parental_traces.append(trace) else: traces_without_parent.append(trace) for trace in parental_traces: trace2 = [copy.copy(i) for i in trace] parent_traces = [trace, trace2] daughter_traces = [i for i in traces_without_parent if i[0].parent in trace] for parent_trace, daughter_trace in izip(parent_traces, daughter_traces): daughter_trace.extend(parent_trace) daughter_trace.sort(key=attrgetter('frame')) return traces_without_parent def extract_division_info_label_id(storage): daughter_cells = [i for i in storage if i.parent is not None] parent_ids = [i.parent_id for i in storage if not np.isnan(i.parent_id)] div_frame = [i.frame for i in storage if not np.isnan(i.parent_id)] return div_frame, daughters_cell_ids, parent_ids def division_frames_and_cell_ids(storage): daughter_cells = [i for i in storage if i.parent is not None] divided_cell_ids = [i.cell_id for i in daughter_cells] div_frame = [i.frame for i in daughter_cells] return div_frame, divided_cell_ids def retrieve_coor(obj): xObj = np.array([i.x for i in obj]).astype(np.float32) yObj = np.array([i.y for i in obj]).astype(np.float32) return xObj, yObj PK!{WBSScelltk/utils/track_utils.pyfrom __future__ import division import numpy as np from scipy.spatial.distance import cdist try: from munkres import munkres except: from _munkres import munkres def calc_ratiodiff(a, b): """calculate how much pairwise ratio of vector a to b. """ a0, a1 = np.meshgrid(a, b) return (a1.T - a0.T)/a0.T def calc_diff(a, b): """calculate how much pairwise ratio of vector a to b. """ a0, a1 = np.meshgrid(a, b) return a1.T - a0.T def calc_massdiff(cells0, cells1): return calc_ratiodiff([i.total_intensity.astype(np.float) for i in cells0], [i.total_intensity.astype(np.float) for i in cells1]) def find_one_to_one_assign(cost): (_, col1) = np.where([np.sum(cost, 0) == 1]) cost[np.sum(cost, 1) != 1] = False (row, col2) = np.where(cost) good_curr_idx = [ci for ri, ci in zip(row, col2) if ci in col1] good_prev_idx = [ri for ri, ci in zip(row, col2) if ci in good_curr_idx] return good_curr_idx, good_prev_idx def prepare_costmat(cost, costDie, costBorn): '''d is cost matrix, often distance matrix where rows are previous and columns as current d contains NaN where tracking of those two objects are not possible. costDie and costBorn''' cost[np.isnan(cost)] = np.Inf # give a large cost. costDieMat = np.array(np.float64(np.diag([costDie]*cost.shape[0]))) # diagonal costBornMat = np.array(np.float64(np.diag([costBorn]*cost.shape[1]))) costDieMat[costDieMat == 0] = np.Inf costBornMat[costBornMat == 0] = np.Inf costMat = np.ones((sum(cost.shape), sum(cost.shape)))*np.Inf costMat[0:cost.shape[0], 0:cost.shape[1]] = cost costMat[-cost.shape[1]:, 0:cost.shape[1]] = costBornMat costMat[0:cost.shape[0], -cost.shape[0]:] = costDieMat lowerRightBlock = cost.transpose() costMat[cost.shape[0]:, cost.shape[1]:] = lowerRightBlock return costMat def call_lap(cost, costDie, costBorn): costMat = prepare_costmat(cost, costDie, costBorn) t = munkres(costMat) topleft = t[0:cost.shape[0], 0:cost.shape[1]] return topleft def pick_closer_binarycostmat(binarymat, distmat): ''' pick closer cells if there are two similar nucleus within area ''' twonuc = np.where(np.sum(binarymat, 1) == 2)[0] for ti in twonuc: di = distmat[ti, :] bi = binarymat[ti, :] binarymat[ti, :] = min(di[bi]) == di return binarymat def pick_closer_cost(binarymat, distmat): distmat = distmat.astype(np.float) bmat = np.zeros(binarymat.shape, np.bool) distmat[~binarymat] = np.Inf true_rows = np.unique(np.where(binarymat)[0]) for i in true_rows: bmat[i, np.argmin(distmat[i, :])] = True return bmat def _find_best_neck_cut(rps0, store, DISPLACEMENT, MASSTHRES): """called by track_neck_cut only """ good_cells = [] for cands in store: if not cands or not rps0: continue dist = cdist([i.centroid for i in rps0], [i.centroid for i in cands]) massdiff = calc_massdiff(rps0, cands) binary_cost = (dist < DISPLACEMENT) * (abs(massdiff) < MASSTHRES) line_int = [i.line_total for i in cands] line_mat = np.tile(line_int, len(rps0)).reshape(len(rps0), len(line_int)) binary_cost = pick_closer_cost(binary_cost, line_mat) binary_cost = pick_closer_cost(binary_cost.T, dist.T).T idx1, idx0 = find_one_to_one_assign(binary_cost.copy()) if not idx0: continue i0, i1 = idx0[0], idx1[0] cell = cands[i1] cell.previous = rps0[i0] good_cells.append(cell) return good_cells def _update_labels_neck_cut(labels0, labels1, good_cells): """called by track_neck_cut only """ labels = -labels1.copy() minint = -np.max(abs(labels)) unique_raw_labels = np.unique([cell.raw_label for cell in good_cells]) neg_labels = np.zeros(labels.shape, np.int32) for i in unique_raw_labels: minint -= 1 neg_labels[labels1 == i] = minint labels[labels1 == i] = 0 for cell in good_cells: for c0, c1 in cell.coords: neg_labels[c0, c1] = cell.previous.label labels0[labels0 == cell.previous.label] = -cell.previous.label labels = labels + neg_labels return labels0, labels def _find_match(rps0, cands, DISPLACEMENT, MASSTHRES): good_cells = [] if not cands or not rps0: return [] dist = cdist([i.centroid for i in rps0], [i.centroid for i in cands]) massdiff = calc_massdiff(rps0, cands) binary_cost = (dist < DISPLACEMENT) * (abs(massdiff) < MASSTHRES) binary_cost = pick_closer_cost(binary_cost, dist) binary_cost = pick_closer_cost(binary_cost.T, dist.T).T idx1, idx0 = find_one_to_one_assign(binary_cost.copy()) if not idx0: return [] i0, i1 = idx0[0], idx1[0] cell = cands[i1] cell.previous = rps0[i0] good_cells.append(cell) return good_cells PK!Uhcelltk/utils/unet_predict.pyfrom __future__ import division import os import numpy as np from os.path import join, basename, splitext from tfutils import imread from tfutils import make_outputdir, normalize from tfutils import pad_image, normalize_predictions import tifffile as tiff import _model_builder def predict(img_path, weight_path): x = imread(img_path) x = normalize(x) if x.ndim == 2: x = np.expand_dims(x, -1) elif x.ndim == 3: x = np.moveaxis(x, 0, -1) x = np.expand_dims(x, 0) num_colors = x.shape[-1] x, hpadding, wpadding = pad_image(x) model = _model_builder.get_model(x.shape[1], x.shape[2], num_colors, activation=None) model.load_weights(weight_path) predictions = model.predict(x, batch_size=1) predictions = [predictions[0, :, :, i] for i in range(predictions.shape[-1])] # resize predictions to match image dimensions (i.e. remove padding) height = np.shape(predictions[0])[0] width = np.shape(predictions[0])[1] predictions = [p[hpadding[0]:height-hpadding[1], wpadding[0]:width-wpadding[1]] for p in predictions] predictions = normalize_predictions(predictions) return predictions def save_output(outputdir, images, pattern): make_outputdir(outputdir) for num, img in enumerate(images): tiff.imsave(join(outputdir, '{0}_l{1}.tif'.format(pattern, num)), img) def _parse_command_line_args(): import argparse parser = argparse.ArgumentParser(description='predict') parser.add_argument('-i', '--image', help='image file path') parser.add_argument('-w', '--weight', help='hdf5 file path') parser.add_argument('-o', '--output', default='.', help='output directory') return parser.parse_args() def _main(): args = _parse_command_line_args() images = predict(args.image, args.weight) save_output(args.output, images, splitext(basename(args.image))[0]) if __name__ == "__main__": _main() PK!ycelltk/utils/util.pyimport numpy as np from scipy.ndimage import imread as imread0 import tifffile as tiff def imread_check_tiff(path): img = imread0(path) if img.dtype == 'object': img = tiff.imread(path) return img def imread(path): if isinstance(path, tuple) or isinstance(path, list): st = [] for p in path: st.append(imread_check_tiff(p)) img = np.dstack(st) if img.shape[2] == 1: np.squeeze(img, axis=2) return img else: return imread_check_tiff(path)PK!##celltk/utils/wavelet_bgr.py#!/usr/bin/python # # Wavelet based background removal as described in this publication: # Galloway et al., An Iterative Algorithm for Background Removal in Spectroscopy by Wavelet Transforms, Applied Spectroscopy, 2009 # # Hazen 01/15 # import numpy import pywt class WaveletBGR(object): def __init__(self, wavelet_type = 'db4', padding_mode = 'sp1'): """ Create a Wavelet background remover. :param wavelet_type: The type of wavelet to use (see the pywt documentation). :type wavelet_type: string. :param padding_mode: How to extrapolate off the ends of the data (see the pywt documentation). :type padding_mode: string. """ self.coeffs = None self.padding_mode = padding_mode self.shape = None self.wavelet_level = None self.wavelet_type = wavelet_type def _estimateBG_(self, image, wavelet_level): """ Private function that estimates the background. """ # Check if requested level is different from the saved level. if (self.wavelet_level is not None) and (self.wavelet_level != wavelet_level): self.coeffs = None self.wavelet_level = wavelet_level # Check if image size is different from the previous level. if (self.shape is not None): if (self.shape[0] != image.shape[0]) or (self.shape[1] != image.shape[1]): self.coeffs = None self.shape = image.shape # 2D Wavelet Transform. coeffs = pywt.wavedec2(image, self.wavelet_type, mode = self.padding_mode, level = wavelet_level) # Set all the details coefficients to zero (if necessary). if self.coeffs is None: self.coeffs = [] self.coeffs.append(coeffs[0]) for i in range(len(coeffs) - 1): temp = [] for j in range(3): temp.append(numpy.zeros(coeffs[i+1][j].shape)) self.coeffs.append(temp) else: self.coeffs[0] = coeffs[0] # 2D Inverse Wavelet Transform. bg_estimate = pywt.waverec2(self.coeffs, self.wavelet_type, mode = self.padding_mode) return bg_estimate def estimateBG(self, image, iterations, threshold, wavelet_level): """ Estimate the background. :param image: The image. :type image: 2D numpy array. :param iterations: Number of iterations to perform to do the estimate. :type iterations: integer. :param threshold: Threshold for maximum difference between the foreground and the background. :type threshold: float or integer. :param wavelet_level: The wavelet level. :type wavelet_level: integer. """ temp = numpy.copy(image) for i in range(iterations): background = self._estimateBG_(temp, wavelet_level) mask = (temp > (background + threshold)) temp[mask] = background[mask] return background def removeBG(self, image, iterations, threshold, wavelet_level): """ Estimate the background and subtract it from the image. :param image: The image. :type image: 2D numpy array. :param iterations: Number of iterations to perform to do the estimate. :type iterations: integer. :param threshold: Threshold for maximum difference between the foreground and the background. :type threshold: float or integer. :param wavelet_level: The wavelet level. :type wavelet_level: integer. """ return image - self.estimateBG(image, iterations, threshold, wavelet_level) if (__name__ == "__main__"): import sys import sa_library.datareader as datareader import sa_library.daxwriter as daxwriter if (len(sys.argv) < 6): print "usage " exit() input_movie = datareader.inferReader(sys.argv[1]) output_dax = daxwriter.DaxWriter("subtracted.dax", 0, 0) iterations = int(sys.argv[4]) threshold = float(sys.argv[5]) wavelet_level = int(sys.argv[3]) offset = 100.0 if (len(sys.argv) == 7): offset = float(sys.argv[6]) wbgr = WaveletBGR(wavelet_type = sys.argv[2]) for i in range(input_movie.filmSize()[2]): if((i%10) == 0): print "Processing frame", i image = input_movie.loadAFrame(i) - offset sub = wbgr.removeBG(image, iterations, threshold, wavelet_level) output_dax.addFrame(sub + offset) output_dax.close() PK!HLq*-%celltk-0.3.dist-info/entry_points.txtN+I/N.,()JN)ɶPzɉ99EVy\\PK! *((celltk-0.3.dist-info/LICENSEMIT License Copyright (c) 2017 braysia Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions: The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software. THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE. 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