celldetective 1.4.1__py3-none-any.whl → 1.4.2__py3-none-any.whl

celldetective/gui/processes/segment_cells.py

@@ -4,8 +4,28 @@ import datetime
 import os
 import json
 import numpy as np
-from celldetective.io import extract_position_name, locate_segmentation_model, auto_load_number_of_frames, load_frames, _check_label_dims, _load_frames_to_segment
-from celldetective.utils import _rescale_labels, _segment_image_with_stardist_model, _segment_image_with_cellpose_model, _prep_stardist_model, _prep_cellpose_model, _get_normalize_kwargs_from_config, extract_experiment_channels, _estimate_scale_factor, _extract_channel_indices_from_config, config_section_to_dict, _extract_nbr_channels_from_config, _get_img_num_per_channel
+from celldetective.io import (
+    extract_position_name,
+    locate_segmentation_model,
+    auto_load_number_of_frames,
+    load_frames,
+    _check_label_dims,
+    _load_frames_to_segment,
+)
+from celldetective.utils import (
+    _rescale_labels,
+    _segment_image_with_stardist_model,
+    _segment_image_with_cellpose_model,
+    _prep_stardist_model,
+    _prep_cellpose_model,
+    _get_normalize_kwargs_from_config,
+    extract_experiment_channels,
+    _estimate_scale_factor,
+    _extract_channel_indices_from_config,
+    config_section_to_dict,
+    _extract_nbr_channels_from_config,
+    _get_img_num_per_channel,
+)
 
 from pathlib import Path, PurePath
 from glob import glob
@@ -13,378 +33,467 @@ from shutil import rmtree
 from tqdm import tqdm
 import numpy as np
 from csbdeep.io import save_tiff_imagej_compatible
-from celldetective.segmentation import segment_frame_from_thresholds, merge_instance_segmentation
+from celldetective.segmentation import (
+    segment_frame_from_thresholds,
+    merge_instance_segmentation,
+)
 import gc
 from art import tprint
 
 import concurrent.futures
 
+
 class BaseSegmentProcess(Process):
 
-	def __init__(self, queue=None, process_args=None, *args, **kwargs):
-		
-		super().__init__(*args, **kwargs)
-		
-		self.queue = queue
+    def __init__(self, queue=None, process_args=None, *args, **kwargs):
+
+        super().__init__(*args, **kwargs)
+
+        self.queue = queue
+
+        if process_args is not None:
+            for key, value in process_args.items():
+                setattr(self, key, value)
 
-		if process_args is not None:
-			for key, value in process_args.items():
-				setattr(self, key, value)
+        tprint("Segment")
 
-		tprint("Segment")
+        # Experiment
+        self.locate_experiment_config()
 
-		# Experiment
-		self.locate_experiment_config()
+        print(f"Position: {extract_position_name(self.pos)}...")
+        print("Configuration file: ", self.config)
+        print(f"Population: {self.mode}...")
+        self.instruction_file = os.sep.join(
+            ["configs", f"segmentation_instructions_{self.mode}.json"]
+        )
 
-		print(f"Position: {extract_position_name(self.pos)}...")
-		print("Configuration file: ",self.config)
-		print(f"Population: {self.mode}...")
-		self.instruction_file = os.sep.join(["configs", f"segmentation_instructions_{self.mode}.json"])
-		
-		self.read_instructions()
-		self.extract_experiment_parameters()
-		self.detect_movie_length()
-		self.write_folders()
-	
-	def read_instructions(self):
-		print('Looking for instruction file...')
-		instr_path = PurePath(self.exp_dir,Path(f"{self.instruction_file}"))
-		if os.path.exists(instr_path):
-			with open(instr_path, 'r') as f:
-				_instructions = json.load(f)
-				print(f"Measurement instruction file successfully loaded...")
-				print(f"Instructions: {_instructions}...")
-			self.flip = _instructions.get("flip", False)
-		else:
-			self.flip = False
+        self.read_instructions()
+        self.extract_experiment_parameters()
+        self.detect_movie_length()
+        self.write_folders()
 
+    def read_instructions(self):
+        print("Looking for instruction file...")
+        instr_path = PurePath(self.exp_dir, Path(f"{self.instruction_file}"))
+        if os.path.exists(instr_path):
+            with open(instr_path, "r") as f:
+                _instructions = json.load(f)
+                print(f"Measurement instruction file successfully loaded...")
+                print(f"Instructions: {_instructions}...")
+            self.flip = _instructions.get("flip", False)
+        else:
+            self.flip = False
 
-	def write_folders(self):
+    def write_folders(self):
 
-		self.mode = self.mode.lower()
-		self.label_folder = f"labels_{self.mode}"
+        self.mode = self.mode.lower()
+        self.label_folder = f"labels_{self.mode}"
 
-		if os.path.exists(self.pos+self.label_folder):
-			print('Erasing the previous labels folder...')
-			rmtree(self.pos+self.label_folder)
-		os.mkdir(self.pos+self.label_folder)
-		print(f'Labels folder successfully generated...')
+        if os.path.exists(self.pos + self.label_folder):
+            print("Erasing the previous labels folder...")
+            rmtree(self.pos + self.label_folder)
+        os.mkdir(self.pos + self.label_folder)
+        print(f"Labels folder successfully generated...")
 
+    def extract_experiment_parameters(self):
 
-	def extract_experiment_parameters(self):
+        self.spatial_calibration = float(
+            config_section_to_dict(self.config, "MovieSettings")["pxtoum"]
+        )
+        self.len_movie = float(
+            config_section_to_dict(self.config, "MovieSettings")["len_movie"]
+        )
+        self.movie_prefix = config_section_to_dict(self.config, "MovieSettings")[
+            "movie_prefix"
+        ]
+        self.nbr_channels = _extract_nbr_channels_from_config(self.config)
+        self.channel_names, self.channel_indices = extract_experiment_channels(
+            self.exp_dir
+        )
 
-		self.spatial_calibration = float(config_section_to_dict(self.config, "MovieSettings")["pxtoum"])
-		self.len_movie = float(config_section_to_dict(self.config, "MovieSettings")["len_movie"])
-		self.movie_prefix = config_section_to_dict(self.config, "MovieSettings")["movie_prefix"]
-		self.nbr_channels = _extract_nbr_channels_from_config(self.config)
-		self.channel_names, self.channel_indices = extract_experiment_channels(self.exp_dir)
+    def locate_experiment_config(self):
 
-	def locate_experiment_config(self):
+        parent1 = Path(self.pos).parent
+        self.exp_dir = parent1.parent
+        self.config = PurePath(self.exp_dir, Path("config.ini"))
 
-		parent1 = Path(self.pos).parent
-		self.exp_dir = parent1.parent
-		self.config = PurePath(self.exp_dir,Path("config.ini"))
+        if not os.path.exists(self.config):
+            print(
+                "The configuration file for the experiment could not be located. Abort."
+            )
+            self.abort_process()
 
-		if not os.path.exists(self.config):
-			print('The configuration file for the experiment could not be located. Abort.')
-			self.abort_process()
+    def detect_movie_length(self):
 
-	def detect_movie_length(self):
+        try:
+            self.file = glob(self.pos + f"movie/{self.movie_prefix}*.tif")[0]
+        except Exception as e:
+            print(f"Error {e}.\nMovie could not be found. Check the prefix.")
+            self.abort_process()
 
-		try:
-			self.file = glob(self.pos+f"movie/{self.movie_prefix}*.tif")[0]
-		except Exception as e:
-			print(f'Error {e}.\nMovie could not be found. Check the prefix.')
-			self.abort_process()
+        len_movie_auto = auto_load_number_of_frames(self.file)
+        if len_movie_auto is not None:
+            self.len_movie = len_movie_auto
 
-		len_movie_auto = auto_load_number_of_frames(self.file)
-		if len_movie_auto is not None:
-			self.len_movie = len_movie_auto
+    def end_process(self):
 
-	def end_process(self):
+        self.terminate()
+        self.queue.put("finished")
 
-		self.terminate()
-		self.queue.put("finished")
+    def abort_process(self):
 
-	def abort_process(self):
-		
-		self.terminate()
-		self.queue.put("error")
+        self.terminate()
+        self.queue.put("error")
 
 
 class SegmentCellDLProcess(BaseSegmentProcess):
-	
-	def __init__(self, *args, **kwargs):
-		
-		super().__init__(*args, **kwargs)
 
-		self.check_gpu()
+    def __init__(self, *args, **kwargs):
+
+        super().__init__(*args, **kwargs)
+
+        self.check_gpu()
+
+        # Model
+        self.locate_model_path()
+        self.extract_model_input_parameters()
+        self.detect_channels()
+        self.detect_rescaling()
 
-		# Model
-		self.locate_model_path()
-		self.extract_model_input_parameters()
-		self.detect_channels()
-		self.detect_rescaling()
+        self.write_log()
 
-		self.write_log()
+        self.sum_done = 0
+        self.t0 = time.time()
 
-		self.sum_done = 0
-		self.t0 = time.time()
+    def extract_model_input_parameters(self):
 
-	def extract_model_input_parameters(self):
-
-		self.required_channels = self.input_config["channels"]
-		if 'selected_channels' in self.input_config:
-			self.required_channels = self.input_config['selected_channels']
-		
-		self.target_cell_size = None
-		if 'target_cell_size_um' in self.input_config and 'cell_size_um' in self.input_config:
-			self.target_cell_size = self.input_config['target_cell_size_um']
-			self.cell_size = self.input_config['cell_size_um']
-
-		self.normalize_kwargs = _get_normalize_kwargs_from_config(self.input_config)
-
-		self.model_type = self.input_config['model_type']
-		self.required_spatial_calibration = self.input_config['spatial_calibration']
-		print(f'Spatial calibration expected by the model: {self.required_spatial_calibration}...')
-		
-		if self.model_type=='cellpose':
-			self.diameter = self.input_config['diameter']
-			self.cellprob_threshold = self.input_config['cellprob_threshold']
-			self.flow_threshold = self.input_config['flow_threshold']
-
-	def write_log(self):
-
-		log=f'segmentation model: {self.model_name}\n'
-		with open(self.pos+f'log_{self.mode}.txt', 'a') as f:
-			f.write(f'{datetime.datetime.now()} SEGMENT \n')
-			f.write(log)
-
-	def detect_channels(self):
-		
-		self.channel_indices = _extract_channel_indices_from_config(self.config, self.required_channels)
-		print(f'Required channels: {self.required_channels} located at channel indices {self.channel_indices}.')
-		self.img_num_channels = _get_img_num_per_channel(self.channel_indices, int(self.len_movie), self.nbr_channels)
-
-	def detect_rescaling(self):
-
-		self.scale = _estimate_scale_factor(self.spatial_calibration, self.required_spatial_calibration)
-		print(f"Scale: {self.scale}...")
-
-		if self.target_cell_size is not None and self.scale is not None:
-			self.scale *= self.cell_size / self.target_cell_size
-		elif self.target_cell_size is not None:
-			if self.target_cell_size != self.cell_size:
-				self.scale = self.cell_size / self.target_cell_size
-
-		print(f"Scale accounting for expected cell size: {self.scale}...")
-
-	def locate_model_path(self):
-
-		self.model_complete_path = locate_segmentation_model(self.model_name)
-		if self.model_complete_path is None:
-			print('Model could not be found. Abort.')
-			self.abort_process()
-		else:
-			print(f'Model path: {self.model_complete_path}...')
-		
-		if not os.path.exists(self.model_complete_path+"config_input.json"):
-			print('The configuration for the inputs to the model could not be located. Abort.')
-			self.abort_process()
-
-		with open(self.model_complete_path+"config_input.json") as config_file:
-			self.input_config = json.load(config_file)
-					
-	def check_gpu(self):
-
-		if not self.use_gpu:
-			os.environ['CUDA_VISIBLE_DEVICES'] = '-1'
-
-	def run(self):
-
-		try:
-
-			if self.model_type=='stardist':
-				model, scale_model = _prep_stardist_model(self.model_name, Path(self.model_complete_path).parent, use_gpu=self.use_gpu, scale=self.scale)
-
-			elif self.model_type=='cellpose':
-				model, scale_model = _prep_cellpose_model(self.model_name, self.model_complete_path, use_gpu=self.use_gpu, n_channels=len(self.required_channels), scale=self.scale)
-
-			list_indices = range(self.len_movie)
-			if self.flip:
-				list_indices = reversed(list_indices)
-
-			for t in tqdm(list_indices,desc="frame"):
-				
-				f = _load_frames_to_segment(self.file, self.img_num_channels[:,t], scale_model=scale_model, normalize_kwargs=self.normalize_kwargs)
-
-				if self.model_type=="stardist":
-					Y_pred = _segment_image_with_stardist_model(f, model=model, return_details=False)
-
-				elif self.model_type=="cellpose":
-					Y_pred = _segment_image_with_cellpose_model(f, model=model, diameter=self.diameter, cellprob_threshold=self.cellprob_threshold, flow_threshold=self.flow_threshold)
-
-				if self.scale is not None:
-					Y_pred = _rescale_labels(Y_pred, scale_model=scale_model)
-				
-				Y_pred = _check_label_dims(Y_pred, file=self.file)
-
-				save_tiff_imagej_compatible(self.pos+os.sep.join([self.label_folder,f"{str(t).zfill(4)}.tif"]), Y_pred, axes='YX')
-				
-				del f;
-				del Y_pred;
-				gc.collect()
-				
-				# Send signal for progress bar
-				self.sum_done+=1/self.len_movie*100
-				mean_exec_per_step = (time.time() - self.t0) / (t+1)
-				pred_time = (self.len_movie - (t+1)) * mean_exec_per_step
-				self.queue.put([self.sum_done, pred_time])
-
-		except Exception as e:
-			print(e)
-
-		try:
-			del model
-		except:
-			pass		
-		
-		gc.collect()
-		print("Done.")
+        self.required_channels = self.input_config["channels"]
+        if "selected_channels" in self.input_config:
+            self.required_channels = self.input_config["selected_channels"]
 
-		# Send end signal
-		self.queue.put("finished")
-		self.queue.close()
+        self.target_cell_size = None
+        if (
+            "target_cell_size_um" in self.input_config
+            and "cell_size_um" in self.input_config
+        ):
+            self.target_cell_size = self.input_config["target_cell_size_um"]
+            self.cell_size = self.input_config["cell_size_um"]
+
+        self.normalize_kwargs = _get_normalize_kwargs_from_config(self.input_config)
+
+        self.model_type = self.input_config["model_type"]
+        self.required_spatial_calibration = self.input_config["spatial_calibration"]
+        print(
+            f"Spatial calibration expected by the model: {self.required_spatial_calibration}..."
+        )
+
+        if self.model_type == "cellpose":
+            self.diameter = self.input_config["diameter"]
+            self.cellprob_threshold = self.input_config["cellprob_threshold"]
+            self.flow_threshold = self.input_config["flow_threshold"]
+
+    def write_log(self):
+
+        log = f"segmentation model: {self.model_name}\n"
+        with open(self.pos + f"log_{self.mode}.txt", "a") as f:
+            f.write(f"{datetime.datetime.now()} SEGMENT \n")
+            f.write(log)
+
+    def detect_channels(self):
+
+        self.channel_indices = _extract_channel_indices_from_config(
+            self.config, self.required_channels
+        )
+        print(
+            f"Required channels: {self.required_channels} located at channel indices {self.channel_indices}."
+        )
+        self.img_num_channels = _get_img_num_per_channel(
+            self.channel_indices, int(self.len_movie), self.nbr_channels
+        )
+
+    def detect_rescaling(self):
+
+        self.scale = _estimate_scale_factor(
+            self.spatial_calibration, self.required_spatial_calibration
+        )
+        print(f"Scale: {self.scale}...")
+
+        if self.target_cell_size is not None and self.scale is not None:
+            self.scale *= self.cell_size / self.target_cell_size
+        elif self.target_cell_size is not None:
+            if self.target_cell_size != self.cell_size:
+                self.scale = self.cell_size / self.target_cell_size
+
+        print(f"Scale accounting for expected cell size: {self.scale}...")
+
+    def locate_model_path(self):
+
+        self.model_complete_path = locate_segmentation_model(self.model_name)
+        if self.model_complete_path is None:
+            print("Model could not be found. Abort.")
+            self.abort_process()
+        else:
+            print(f"Model path: {self.model_complete_path}...")
+
+        if not os.path.exists(self.model_complete_path + "config_input.json"):
+            print(
+                "The configuration for the inputs to the model could not be located. Abort."
+            )
+            self.abort_process()
+
+        with open(self.model_complete_path + "config_input.json") as config_file:
+            self.input_config = json.load(config_file)
+
+    def check_gpu(self):
+
+        if not self.use_gpu:
+            os.environ["CUDA_VISIBLE_DEVICES"] = "-1"
+
+    def run(self):
+
+        try:
+
+            if self.model_type == "stardist":
+                model, scale_model = _prep_stardist_model(
+                    self.model_name,
+                    Path(self.model_complete_path).parent,
+                    use_gpu=self.use_gpu,
+                    scale=self.scale,
+                )
+
+            elif self.model_type == "cellpose":
+                model, scale_model = _prep_cellpose_model(
+                    self.model_name,
+                    self.model_complete_path,
+                    use_gpu=self.use_gpu,
+                    n_channels=len(self.required_channels),
+                    scale=self.scale,
+                )
+
+            list_indices = range(self.len_movie)
+            if self.flip:
+                list_indices = reversed(list_indices)
+
+            for i, t in enumerate(tqdm(list_indices, desc="frame")):
+
+                f = _load_frames_to_segment(
+                    self.file,
+                    self.img_num_channels[:, t],
+                    scale_model=scale_model,
+                    normalize_kwargs=self.normalize_kwargs,
+                )
+
+                if self.model_type == "stardist":
+                    Y_pred = _segment_image_with_stardist_model(
+                        f, model=model, return_details=False
+                    )
+
+                elif self.model_type == "cellpose":
+                    Y_pred = _segment_image_with_cellpose_model(
+                        f,
+                        model=model,
+                        diameter=self.diameter,
+                        cellprob_threshold=self.cellprob_threshold,
+                        flow_threshold=self.flow_threshold,
+                    )
+
+                if self.scale is not None:
+                    Y_pred = _rescale_labels(Y_pred, scale_model=scale_model)
+
+                Y_pred = _check_label_dims(Y_pred, file=self.file)
+
+                save_tiff_imagej_compatible(
+                    self.pos
+                    + os.sep.join([self.label_folder, f"{str(t).zfill(4)}.tif"]),
+                    Y_pred,
+                    axes="YX",
+                )
+
+                del f
+                del Y_pred
+                gc.collect()
+
+                # Send signal for progress bar
+                self.sum_done += 1 / self.len_movie * 100
+                mean_exec_per_step = (time.time() - self.t0) / (i + 1)
+                pred_time = (self.len_movie - (i + 1)) * mean_exec_per_step
+                self.queue.put([self.sum_done, pred_time])
+
+        except Exception as e:
+            print(e)
+
+        try:
+            del model
+        except:
+            pass
+
+        gc.collect()
+        print("Done.")
+
+        # Send end signal
+        self.queue.put("finished")
+        self.queue.close()
 
 
 class SegmentCellThresholdProcess(BaseSegmentProcess):
-	
-	def __init__(self, *args, **kwargs):
-		
-		super().__init__(*args, **kwargs)
 
-		self.equalize = False
+    def __init__(self, *args, **kwargs):
+
+        super().__init__(*args, **kwargs)
+
+        self.equalize = False
+
+        # Model
+
+        self.load_threshold_config()
+        self.extract_threshold_parameters()
+        self.detect_channels()
+        self.prepare_equalize()
 
-		# Model
+        self.write_log()
 
-		self.load_threshold_config()
-		self.extract_threshold_parameters()
-		self.detect_channels()
-		self.prepare_equalize()
+        self.sum_done = 0
+        self.t0 = time.time()
 
-		self.write_log()
+    def prepare_equalize(self):
 
-		self.sum_done = 0
-		self.t0 = time.time()
+        for i in range(len(self.instructions)):
 
-	def prepare_equalize(self):
+            if self.equalize[i]:
+                f_reference = load_frames(
+                    self.img_num_channels[:, self.equalize_time[i]],
+                    self.file,
+                    scale=None,
+                    normalize_input=False,
+                )
+                f_reference = f_reference[:, :, self.instructions[i]["target_channel"]]
+            else:
+                f_reference = None
+
+            self.instructions[i].update({"equalize_reference": f_reference})
+
+    def load_threshold_config(self):
 
-		for i in range(len(self.instructions)):
+        self.instructions = []
+        for inst in self.threshold_instructions:
+            if os.path.exists(inst):
+                with open(inst, "r") as f:
+                    self.instructions.append(json.load(f))
+            else:
+                print("The configuration path is not valid. Abort.")
+                self.abort_process()
 
-			if self.equalize[i]:
-				f_reference = load_frames(self.img_num_channels[:,self.equalize_time[i]], self.file, scale=None, normalize_input=False)
-				f_reference = f_reference[:,:,self.instructions[i]['target_channel']]
-			else:
-				f_reference = None
-
-			self.instructions[i].update({'equalize_reference': f_reference})
-
-	def load_threshold_config(self):
-
-		self.instructions = []
-		for inst in self.threshold_instructions:
-			if os.path.exists(inst):
-				with open(inst, 'r') as f:
-					self.instructions.append(json.load(f))
-			else:
-				print('The configuration path is not valid. Abort.')
-				self.abort_process()
-
-	def extract_threshold_parameters(self):
-		
-		self.required_channels = []
-		self.equalize = []
-		self.equalize_time = []
-
-		for i in range(len(self.instructions)):
-			ch = [self.instructions[i]['target_channel']]
-			self.required_channels.append(ch)
-
-			if 'equalize_reference' in self.instructions[i]:
-				equalize, equalize_time = self.instructions[i]['equalize_reference']	
-				self.equalize.append(equalize)
-				self.equalize_time.append(equalize_time)
-
-	def write_log(self):
-
-		log=f'Threshold segmentation: {self.threshold_instructions}\n'
-		with open(self.pos+f'log_{self.mode}.txt', 'a') as f:
-			f.write(f'{datetime.datetime.now()} SEGMENT \n')
-			f.write(log)
-
-	def detect_channels(self):
-
-		for i in range(len(self.instructions)):
-			
-			self.channel_indices = _extract_channel_indices_from_config(self.config, self.required_channels[i])
-			print(f'Required channels: {self.required_channels[i]} located at channel indices {self.channel_indices}.')
-			self.instructions[i].update({'target_channel': self.channel_indices[0]})
-			self.instructions[i].update({'channel_names': self.channel_names})
-		
-		self.img_num_channels = _get_img_num_per_channel(np.arange(self.nbr_channels), self.len_movie, self.nbr_channels)
-
-	def parallel_job(self, indices):
-
-		try:
-
-			for t in tqdm(indices,desc="frame"): #for t in tqdm(range(self.len_movie),desc="frame"):
-				
-				# Load channels at time t
-				masks = []
-				for i in range(len(self.instructions)):
-					f = load_frames(self.img_num_channels[:,t], self.file, scale=None, normalize_input=False)
-					mask = segment_frame_from_thresholds(f, **self.instructions[i])
-					#print(f'Frame {t}; segment with {self.instructions[i]=}...')
-					masks.append(mask)
-
-				if len(self.instructions)>1:
-					mask = merge_instance_segmentation(masks, mode='OR')
-
-				save_tiff_imagej_compatible(os.sep.join([self.pos, self.label_folder, f"{str(t).zfill(4)}.tif"]), mask.astype(np.uint16), axes='YX')
-
-				del f;
-				del mask;
-				gc.collect()
-
-				# Send signal for progress bar
-				self.sum_done+=1/self.len_movie*100
-				mean_exec_per_step = (time.time() - self.t0) / (self.sum_done*self.len_movie / 100 + 1)
-				pred_time = (self.len_movie - (self.sum_done*self.len_movie / 100 + 1)) * mean_exec_per_step
-				self.queue.put([self.sum_done, pred_time])
+    def extract_threshold_parameters(self):
 
-		except Exception as e:
-			print(e)
-
-		return
-			
-
-	def run(self):
-
-		self.indices = list(range(self.img_num_channels.shape[1]))
-		if self.flip:
-			self.indices = np.array(list(reversed(self.indices)))
-		
-		chunks = np.array_split(self.indices, self.n_threads)
-
-		with concurrent.futures.ThreadPoolExecutor(max_workers=self.n_threads) as executor:
-			results = results = executor.map(self.parallel_job, chunks) #list(map(lambda x: executor.submit(self.parallel_job, x), chunks))
-			try:
-				for i,return_value in enumerate(results):
-					print(f"Thread {i} output check: ",return_value)
-			except Exception as e:
-				print("Exception: ", e)
-
-		print('Done.')
-		# Send end signal
-		self.queue.put("finished")
-		self.queue.close()
+        self.required_channels = []
+        self.equalize = []
+        self.equalize_time = []
+
+        for i in range(len(self.instructions)):
+            ch = [self.instructions[i]["target_channel"]]
+            self.required_channels.append(ch)
+
+            if "equalize_reference" in self.instructions[i]:
+                equalize, equalize_time = self.instructions[i]["equalize_reference"]
+                self.equalize.append(equalize)
+                self.equalize_time.append(equalize_time)
+
+    def write_log(self):
+
+        log = f"Threshold segmentation: {self.threshold_instructions}\n"
+        with open(self.pos + f"log_{self.mode}.txt", "a") as f:
+            f.write(f"{datetime.datetime.now()} SEGMENT \n")
+            f.write(log)
+
+    def detect_channels(self):
+
+        for i in range(len(self.instructions)):
+
+            self.channel_indices = _extract_channel_indices_from_config(
+                self.config, self.required_channels[i]
+            )
+            print(
+                f"Required channels: {self.required_channels[i]} located at channel indices {self.channel_indices}."
+            )
+            self.instructions[i].update({"target_channel": self.channel_indices[0]})
+            self.instructions[i].update({"channel_names": self.channel_names})
+
+        self.img_num_channels = _get_img_num_per_channel(
+            np.arange(self.nbr_channels), self.len_movie, self.nbr_channels
+        )
+
+    def parallel_job(self, indices):
+
+        try:
+
+            for t in tqdm(
+                indices, desc="frame"
+            ):  # for t in tqdm(range(self.len_movie),desc="frame"):
+
+                # Load channels at time t
+                masks = []
+                for i in range(len(self.instructions)):
+                    f = load_frames(
+                        self.img_num_channels[:, t],
+                        self.file,
+                        scale=None,
+                        normalize_input=False,
+                    )
+                    mask = segment_frame_from_thresholds(f, **self.instructions[i])
+                    # print(f'Frame {t}; segment with {self.instructions[i]=}...')
+                    masks.append(mask)
+
+                if len(self.instructions) > 1:
+                    mask = merge_instance_segmentation(masks, mode="OR")
+
+                save_tiff_imagej_compatible(
+                    os.sep.join(
+                        [self.pos, self.label_folder, f"{str(t).zfill(4)}.tif"]
+                    ),
+                    mask.astype(np.uint16),
+                    axes="YX",
+                )
+
+                del f
+                del mask
+                gc.collect()
+
+                # Send signal for progress bar
+                self.sum_done += 1 / self.len_movie * 100
+                mean_exec_per_step = (time.time() - self.t0) / (
+                    self.sum_done * self.len_movie / 100 + 1
+                )
+                pred_time = (
+                    self.len_movie - (self.sum_done * self.len_movie / 100 + 1)
+                ) * mean_exec_per_step
+                self.queue.put([self.sum_done, pred_time])
+
+        except Exception as e:
+            print(e)
+
+        return
+
+    def run(self):
+
+        self.indices = list(range(self.img_num_channels.shape[1]))
+        if self.flip:
+            self.indices = np.array(list(reversed(self.indices)))
+
+        chunks = np.array_split(self.indices, self.n_threads)
+
+        with concurrent.futures.ThreadPoolExecutor(
+            max_workers=self.n_threads
+        ) as executor:
+            results = results = executor.map(
+                self.parallel_job, chunks
+            )  # list(map(lambda x: executor.submit(self.parallel_job, x), chunks))
+            try:
+                for i, return_value in enumerate(results):
+                    print(f"Thread {i} output check: ", return_value)
+            except Exception as e:
+                print("Exception: ", e)
+
+        print("Done.")
+        # Send end signal
+        self.queue.put("finished")
+        self.queue.close()