def segment_images(inpDir, outDir, config_data): """ Workflow for data with similar morphology as connexin-43 Args: inpDir : path to the input directory outDir : path to the output directory config_data : path to the configuration file """ logging.basicConfig( format='%(asctime)s - %(name)-8s - %(levelname)-8s - %(message)s', datefmt='%d-%b-%y %H:%M:%S') logger = logging.getLogger("main") logger.setLevel(logging.INFO) inpDir_files = os.listdir(inpDir) for i, f in enumerate(inpDir_files): logger.info('Segmenting image : {}'.format(f)) # Load image br = BioReader(os.path.join(inpDir, f)) image = br.read_image() structure_channel = 0 struct_img0 = image[:, :, :, structure_channel, 0] struct_img0 = struct_img0.transpose(2, 0, 1).astype(np.float32) # main algorithm intensity_scaling_param = config_data['intensity_scaling_param'] struct_img = intensity_normalization( struct_img0, scaling_param=intensity_scaling_param) gaussian_smoothing_sigma = config_data['gaussian_smoothing_sigma'] structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma) s3_param = config_data['s3_param'] bw = dot_3d_wrapper(structure_img_smooth, s3_param) minArea = config_data['minArea'] seg = remove_small_objects(bw > 0, min_size=minArea, connectivity=1, in_place=False) seg = seg > 0 out_img = seg.astype(np.uint8) out_img[out_img > 0] = 255 # create output image out_img = out_img.transpose(1, 2, 0) out_img = out_img.reshape( (out_img.shape[0], out_img.shape[1], out_img.shape[2], 1, 1)) # write image using BFIO bw = BioWriter(os.path.join(outDir, f), metadata=br.read_metadata()) bw.num_x(out_img.shape[1]) bw.num_y(out_img.shape[0]) bw.num_z(out_img.shape[2]) bw.num_c(out_img.shape[3]) bw.num_t(out_img.shape[4]) bw.pixel_type(dtype='uint8') bw.write_image(out_img) bw.close_image()
def Workflow_slc25a17( struct_img: np.ndarray, rescale_ratio: float = -1, output_type: str = "default", output_path: Union[str, Path] = None, fn: Union[str, Path] = None, output_func=None, ): """ classic segmentation workflow wrapper for structure SLC25A17 Parameter: ----------- struct_img: np.ndarray the 3D image to be segmented rescale_ratio: float an optional parameter to allow rescale the image before running the segmentation functions, default is no rescaling output_type: str select how to handle output. Currently, four types are supported: 1. default: the result will be saved at output_path whose filename is original name without extention + "_struct_segmentaiton.tiff" 2. array: the segmentation result will be simply returned as a numpy array 3. array_with_contour: segmentation result will be returned together with the contour of the segmentation 4. customize: pass in an extra output_func to do a special save. All the intermediate results, names of these results, the output_path, and the original filename (without extension) will be passed in to output_func. """ ########################################################################## # PARAMETERS: # note that these parameters are supposed to be fixed for the structure # and work well accross different datasets intensity_norm_param = [2, 36] gaussian_smoothing_sigma = 1 gaussian_smoothing_truncate_range = 3.0 dot_3d_sigma = 1 dot_3d_cutoff = 0.045 # 0.03 #0.04 minArea = 3 # 5 ########################################################################## out_img_list = [] out_name_list = [] ################### # PRE_PROCESSING ################### # intenisty normalization (min/max) struct_img = intensity_normalization(struct_img, scaling_param=intensity_norm_param) out_img_list.append(struct_img.copy()) out_name_list.append("im_norm") # rescale if needed if rescale_ratio > 0: struct_img = zoom(struct_img, (1, rescale_ratio, rescale_ratio), order=2) struct_img = (struct_img - struct_img.min() + 1e-8) / (struct_img.max() - struct_img.min() + 1e-8) gaussian_smoothing_truncate_range = ( gaussian_smoothing_truncate_range * rescale_ratio) # smoothing with gaussian filter structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma, truncate_range=gaussian_smoothing_truncate_range, ) out_img_list.append(structure_img_smooth.copy()) out_name_list.append("im_smooth") ################### # core algorithm ################### # step 1: LOG 3d response = dot_3d(structure_img_smooth, log_sigma=dot_3d_sigma) bw = response > dot_3d_cutoff bw = remove_small_objects(bw > 0, min_size=minArea, connectivity=1, in_place=False) out_img_list.append(bw.copy()) out_name_list.append("interm_mask") # step 2: 'local_maxi + watershed' for cell cutting local_maxi = peak_local_max(struct_img, labels=label(bw), min_distance=2, indices=False) out_img_list.append(local_maxi.copy()) out_name_list.append("interm_local_max") distance = distance_transform_edt(bw) im_watershed = watershed( -1 * distance, label(dilation(local_maxi, selem=ball(1))), mask=bw, watershed_line=True, ) ################### # POST-PROCESSING ################### seg = remove_small_objects(im_watershed, min_size=minArea, connectivity=1, in_place=False) # HACK: Only for 2019 April Release ##### if np.count_nonzero(seg > 0) < 50000: print("FLAG: please check the meta data of the original CZI for QC") # output seg = seg > 0 seg = seg.astype(np.uint8) seg[seg > 0] = 255 out_img_list.append(seg.copy()) out_name_list.append("bw_final") if output_type == "default": # the default final output, simply save it to the output path save_segmentation(seg, False, Path(output_path), fn) elif output_type == "customize": # the hook for passing in a customized output function # use "out_img_list" and "out_name_list" in your hook to # customize your output functions output_func(out_img_list, out_name_list, Path(output_path), fn) elif output_type == "array": return seg elif output_type == "array_with_contour": return (seg, generate_segmentation_contour(seg)) else: raise NotImplementedError("invalid output type: {output_type}")
def Workflow_lamp1( struct_img: np.ndarray, rescale_ratio: float = -1, output_type: str = "default", output_path: Union[str, Path] = None, fn: Union[str, Path] = None, output_func=None, ): """ classic segmentation workflow wrapper for structure LAMP1 Parameter: ----------- struct_img: np.ndarray the 3D image to be segmented rescale_ratio: float an optional parameter to allow rescale the image before running the segmentation functions, default is no rescaling output_type: str select how to handle output. Currently, four types are supported: 1. default: the result will be saved at output_path whose filename is original name without extention + "_struct_segmentaiton.tiff" 2. array: the segmentation result will be simply returned as a numpy array 3. array_with_contour: segmentation result will be returned together with the contour of the segmentation 4. customize: pass in an extra output_func to do a special save. All the intermediate results, names of these results, the output_path, and the original filename (without extension) will be passed in to output_func. """ ########################################################################## # PARAMETERS: # note that these parameters are supposed to be fixed for the structure # and work well accross different datasets intensity_scaling_param = [3, 19] gaussian_smoothing_sigma = 1 gaussian_smoothing_truncate_range = 3.0 minArea = 15 # ves_th_2d = 0.1 vesselness_sigma = [1] vesselness_cutoff = 0.15 # hole_min = 60 hole_max = 1600 log_sigma_1 = 5 log_cutoff_1 = 0.09 log_sigma_2 = 2.5 log_cutoff_2 = 0.07 log_sigma_3 = 1 log_cutoff_3 = 0.01 ########################################################################## out_img_list = [] out_name_list = [] # intenisty normalization struct_img = intensity_normalization( struct_img, scaling_param=intensity_scaling_param ) out_img_list.append(struct_img.copy()) out_name_list.append("im_norm") if rescale_ratio > 0: struct_img = zoom(struct_img, (1, rescale_ratio, rescale_ratio), order=2) struct_img = (struct_img - struct_img.min() + 1e-8) / ( struct_img.max() - struct_img.min() + 1e-8 ) gaussian_smoothing_truncate_range = ( gaussian_smoothing_truncate_range * rescale_ratio ) structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma, truncate_range=gaussian_smoothing_truncate_range, ) out_img_list.append(structure_img_smooth.copy()) out_name_list.append("im_smooth") # spot detection response1 = dot_slice_by_slice(structure_img_smooth, log_sigma=log_sigma_1) bw1 = response1 > log_cutoff_1 response2 = dot_slice_by_slice(structure_img_smooth, log_sigma=log_sigma_2) bw2 = response2 > log_cutoff_2 bw_spot = np.logical_or(bw1, bw2) response3 = dot_slice_by_slice(structure_img_smooth, log_sigma=log_sigma_3) bw3 = response3 > log_cutoff_3 bw_spot = np.logical_or(bw_spot, bw3) # ring/filament detection ves = vesselnessSliceBySlice( structure_img_smooth, sigmas=vesselness_sigma, tau=1, whiteonblack=True ) bw_ves = ves > vesselness_cutoff # fill holes partial_fill = np.logical_or(bw_spot, bw_ves) out_img_list.append(partial_fill.copy()) out_name_list.append("interm_before_hole") holes = np.zeros_like(partial_fill) for zz in range(partial_fill.shape[0]): background_lab = label(~partial_fill[zz, :, :], connectivity=1) out = np.copy(background_lab) component_sizes = np.bincount(background_lab.ravel()) too_big = component_sizes > hole_max too_big_mask = too_big[background_lab] out[too_big_mask] = 0 # too_small = component_sizes <hole_min # too_small_mask = too_small[background_lab] # out[too_small_mask] = 0 holes[zz, :, :] = out full_fill = np.logical_or(partial_fill, holes) seg = remove_small_objects( full_fill, min_size=minArea, connectivity=1, in_place=False ) # output seg = seg > 0 seg = seg.astype(np.uint8) seg[seg > 0] = 255 out_img_list.append(seg.copy()) out_name_list.append("bw_final") if output_type == "default": # the default final output, simply save it to the output path save_segmentation(seg, False, Path(output_path), fn) elif output_type == "customize": # the hook for passing in a customized output function # use "out_img_list" and "out_name_list" in your hook to # customize your output functions output_func(out_img_list, out_name_list, Path(output_path), fn) elif output_type == "array": return seg elif output_type == "array_with_contour": return (seg, generate_segmentation_contour(seg)) else: raise NotImplementedError("invalid output type: {output_type}")
def Workflow_rab5a( struct_img: np.ndarray, rescale_ratio: float = -1, output_type: str = "default", output_path: Union[str, Path] = None, fn: Union[str, Path] = None, output_func=None, ): """ classic segmentation workflow wrapper for structure RAB5A Parameter: ----------- struct_img: np.ndarray the 3D image to be segmented rescale_ratio: float an optional parameter to allow rescale the image before running the segmentation functions, default is no rescaling output_type: str select how to handle output. Currently, four types are supported: 1. default: the result will be saved at output_path whose filename is original name without extention + "_struct_segmentaiton.tiff" 2. array: the segmentation result will be simply returned as a numpy array 3. array_with_contour: segmentation result will be returned together with the contour of the segmentation 4. customize: pass in an extra output_func to do a special save. All the intermediate results, names of these results, the output_path, and the original filename (without extension) will be passed in to output_func. """ ########################################################################## # PARAMETERS: # note that these parameters are supposed to be fixed for the structure # and work well accross different datasets intensity_norm_param = [1000] gaussian_smoothing_sigma = 1 gaussian_smoothing_truncate_range = 3.0 dot_3d_sigma = 1 dot_3d_cutoff = 0.03 hole_min = 0 hole_max = 81 minArea = 3 ########################################################################## out_img_list = [] out_name_list = [] ################### # PRE_PROCESSING ################### # intenisty normalization (min/max) struct_img = intensity_normalization(struct_img, scaling_param=intensity_norm_param) out_img_list.append(struct_img.copy()) out_name_list.append("im_norm") # rescale if needed if rescale_ratio > 0: struct_img = zoom(struct_img, (1, rescale_ratio, rescale_ratio), order=2) struct_img = (struct_img - struct_img.min() + 1e-8) / (struct_img.max() - struct_img.min() + 1e-8) gaussian_smoothing_truncate_range = ( gaussian_smoothing_truncate_range * rescale_ratio) # smoothing with gaussian filter structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma, truncate_range=gaussian_smoothing_truncate_range, ) out_img_list.append(structure_img_smooth.copy()) out_name_list.append("im_smooth") ################### # core algorithm ################### # step 1: LOG 3d response = dot_3d(structure_img_smooth, log_sigma=dot_3d_sigma) bw = response > dot_3d_cutoff # step 2: fill holes and remove small objects bw_filled = hole_filling(bw, hole_min, hole_max, True) seg = remove_small_objects(bw_filled, min_size=minArea, connectivity=1, in_place=False) # output seg = seg > 0 seg = seg.astype(np.uint8) seg[seg > 0] = 255 out_img_list.append(seg.copy()) out_name_list.append("bw_final") if output_type == "default": # the default final output, simply save it to the output path save_segmentation(seg, False, Path(output_path), fn) elif output_type == "customize": # the hook for passing in a customized output function # use "out_img_list" and "out_name_list" in your hook to # customize your output functions output_func(out_img_list, out_name_list, Path(output_path), fn) elif output_type == "array": return seg elif output_type == "array_with_contour": return (seg, generate_segmentation_contour(seg)) else: raise NotImplementedError("invalid output type: {output_type}")
def segment_images(inpDir, outDir, config_data): """ Workflow for dot like shapes such as Centrin-2, Desmoplakin, PMP34. Args: inpDir : path to the input directory outDir : path to the output directory config_data : path to the configuration file """ logging.basicConfig( format='%(asctime)s - %(name)-8s - %(levelname)-8s - %(message)s', datefmt='%d-%b-%y %H:%M:%S') logger = logging.getLogger("main") logger.setLevel(logging.INFO) inpDir_files = os.listdir(inpDir) for i, f in enumerate(inpDir_files): logger.info('Segmenting image : {}'.format(f)) # Load an image br = BioReader(os.path.join(inpDir, f)) image = br.read_image() structure_channel = 0 struct_img0 = image[:, :, :, structure_channel, 0] struct_img0 = struct_img0.transpose(2, 0, 1).astype(np.float32) # main algorithm intensity_scaling_param = config_data['intensity_scaling_param'] struct_img = intensity_normalization( struct_img0, scaling_param=intensity_scaling_param) gaussian_smoothing_sigma = config_data['gaussian_smoothing_sigma'] if config_data["gaussian_smoothing"] == "gaussian_slice_by_slice": structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma) else: structure_img_smooth = image_smoothing_gaussian_3d( struct_img, sigma=gaussian_smoothing_sigma) s3_param = config_data['s3_param'] bw = dot_3d_wrapper(structure_img_smooth, s3_param) minArea = config_data['minArea'] Mask = remove_small_objects(bw > 0, min_size=minArea, connectivity=1, in_place=False) Seed = dilation(peak_local_max(struct_img, labels=label(Mask), min_distance=2, indices=False), selem=ball(1)) Watershed_Map = -1 * distance_transform_edt(bw) seg = watershed(Watershed_Map, label(Seed), mask=Mask, watershed_line=True) seg = remove_small_objects(seg > 0, min_size=minArea, connectivity=1, in_place=False) seg = seg > 0 out_img = seg.astype(np.uint8) out_img[out_img > 0] = 255 # create output image out_img = out_img.transpose(1, 2, 0) out_img = out_img.reshape( (out_img.shape[0], out_img.shape[1], out_img.shape[2], 1, 1)) # write image using BFIO bw = BioWriter(os.path.join(outDir, f)) bw.num_x(out_img.shape[1]) bw.num_y(out_img.shape[0]) bw.num_z(out_img.shape[2]) bw.num_c(out_img.shape[3]) bw.num_t(out_img.shape[4]) bw.pixel_type(dtype='uint8') bw.write_image(out_img) bw.close_image()
def FABP3_Cardio_Pipeline(struct_img, rescale_ratio, output_type, output_path, fn, output_func=None): ########################################################################## # PARAMETERS: # note that these parameters are supposed to be fixed for the structure # and work well accross different datasets # ADD you parameters here intensity_norm_param = [1, 18] minArea = 0 gaussian_smoothing_sigma = 1 # lower_bound = 685 # upper_b = 18 # ADD-HERE ########################################################################## out_img_list = [] out_name_list = [] ################### # PRE_PROCESSING ################### # intenisty normalization (min/max) struct_img = intensity_normalization(struct_img, scaling_param=intensity_norm_param) # upper_bound = np.mean(struct_img) + 18 * np.std(struct_img) # struct_img[struct_img < lower_bound] = lower_bound # struct_img[struct_img > upper_bound] = upper_bound # struct_img = (struct_img - lower_bound).astype(float)/(upper_bound - lower_bound) out_img_list.append(struct_img.copy()) out_name_list.append("im_norm") # rescale if needed if rescale_ratio > 0: struct_img = resize(struct_img, [1, rescale_ratio, rescale_ratio], method="cubic") struct_img = (struct_img - struct_img.min() + 1e-8) / (struct_img.max() - struct_img.min() + 1e-8) # smoothing with gaussian filter structure_img_smooth = image_smoothing_gaussian_slice_by_slice( struct_img, sigma=gaussian_smoothing_sigma) # ADD-HERE out_img_list.append(structure_img_smooth.copy()) out_name_list.append("im_smooth") # core algorithm # th_li = threshold_li(structure_img_smooth) # PARAMETERS for this step ## s2_param = [[1, 0.02]] bw = dot_2d_slice_by_slice_wrapper(structure_img_smooth, s2_param) # POST-PROCESSING seg = remove_small_objects(bw, min_size=minArea, connectivity=1, in_place=False) # output seg = seg > 0 seg = seg.astype(np.uint8) seg[seg > 0] = 255 out_img_list.append(seg.copy()) out_name_list.append("bw_final") if output_type == "default": # the default final output save_segmentation(seg, False, output_path, fn) elif output_type == "TEMPLATE": # the hook for pre-defined output functions template_output(out_img_list, out_name_list, output_type, output_path, fn)