def get_skeleton_points(self, obj): '''Get points by skeletonizing the objects and decimating''' ii = [] jj = [] total_skel = np.zeros(obj.shape, bool) for labels, indexes in obj.get_labels(): colors = morph.color_labels(labels) for color in range(1, np.max(colors) + 1): labels_mask = colors == color skel = morph.skeletonize( labels_mask, ordering=distance_transform_edt(labels_mask) * poisson_equation(labels_mask)) total_skel = total_skel | skel n_pts = np.sum(total_skel) if n_pts == 0: return np.zeros(0, np.int32), np.zeros(0, np.int32) i, j = np.where(total_skel) if n_pts > self.max_points.value: # # Decimate the skeleton by finding the branchpoints in the # skeleton and propagating from those. # markers = np.zeros(total_skel.shape, np.int32) branchpoints = \ morph.branchpoints(total_skel) | morph.endpoints(total_skel) markers[branchpoints] = np.arange(np.sum(branchpoints)) + 1 # # We compute the propagation distance to that point, then impose # a slightly arbitarary order to get an unambiguous ordering # which should number the pixels in a skeleton branch monotonically # ts_labels, distances = propagate(np.zeros(markers.shape), markers, total_skel, 1) order = np.lexsort((j, i, distances[i, j], ts_labels[i, j])) # # Get a linear space of self.max_points elements with bounds at # 0 and len(order)-1 and use that to select the points. # order = order[np.linspace(0, len(order) - 1, self.max_points.value).astype(int)] return i[order], j[order] return i, j
def get_skeleton_points(self, obj): '''Get points by skeletonizing the objects and decimating''' ii = [] jj = [] total_skel = np.zeros(obj.shape, bool) for labels, indexes in obj.get_labels(): colors = morph.color_labels(labels) for color in range(1, np.max(colors) + 1): labels_mask = colors == color skel = morph.skeletonize( labels_mask, ordering = distance_transform_edt(labels_mask) * poisson_equation(labels_mask)) total_skel = total_skel | skel n_pts = np.sum(total_skel) if n_pts == 0: return np.zeros(0, np.int32), np.zeros(0, np.int32) i, j = np.where(total_skel) if n_pts > self.max_points.value: # # Decimate the skeleton by finding the branchpoints in the # skeleton and propagating from those. # markers = np.zeros(total_skel.shape, np.int32) branchpoints = \ morph.branchpoints(total_skel) | morph.endpoints(total_skel) markers[branchpoints] = np.arange(np.sum(branchpoints))+1 # # We compute the propagation distance to that point, then impose # a slightly arbitarary order to get an unambiguous ordering # which should number the pixels in a skeleton branch monotonically # ts_labels, distances = propagate(np.zeros(markers.shape), markers, total_skel, 1) order = np.lexsort((j, i, distances[i, j], ts_labels[i, j])) # # Get a linear space of self.max_points elements with bounds at # 0 and len(order)-1 and use that to select the points. # order = order[ np.linspace(0, len(order)-1, self.max_points.value).astype(int)] return i[order], j[order] return i, j
def do_measurements(self, workspace, image_name, object_name, center_object_name, center_choice, bin_count_settings, dd): '''Perform the radial measurements on the image set workspace - workspace that holds images / objects image_name - make measurements on this image object_name - make measurements on these objects center_object_name - use the centers of these related objects as the centers for radial measurements. None to use the objects themselves. center_choice - the user's center choice for this object: C_SELF, C_CENTERS_OF_OBJECTS or C_EDGES_OF_OBJECTS. bin_count_settings - the bin count settings group d - a dictionary for saving reusable partial results returns one statistics tuple per ring. ''' assert isinstance(workspace, cpw.Workspace) assert isinstance(workspace.object_set, cpo.ObjectSet) bin_count = bin_count_settings.bin_count.value wants_scaled = bin_count_settings.wants_scaled.value maximum_radius = bin_count_settings.maximum_radius.value image = workspace.image_set.get_image(image_name, must_be_grayscale=True) objects = workspace.object_set.get_objects(object_name) labels, pixel_data = cpo.crop_labels_and_image(objects.segmented, image.pixel_data) nobjects = np.max(objects.segmented) measurements = workspace.measurements assert isinstance(measurements, cpmeas.Measurements) heatmaps = {} for heatmap in self.heatmaps: if heatmap.object_name.get_objects_name() == object_name and \ image_name == heatmap.image_name.get_image_name() and \ heatmap.get_number_of_bins() == bin_count: dd[id(heatmap)] = \ heatmaps[MEASUREMENT_ALIASES[heatmap.measurement.value]] = \ np.zeros(labels.shape) if nobjects == 0: for bin in range(1, bin_count + 1): for feature in (F_FRAC_AT_D, F_MEAN_FRAC, F_RADIAL_CV): feature_name = ( (feature + FF_GENERIC) % (image_name, bin, bin_count)) measurements.add_measurement( object_name, "_".join([M_CATEGORY, feature_name]), np.zeros(0)) if not wants_scaled: measurement_name = "_".join([M_CATEGORY, feature, image_name, FF_OVERFLOW]) measurements.add_measurement( object_name, measurement_name, np.zeros(0)) return [(image_name, object_name, "no objects", "-", "-", "-", "-")] name = (object_name if center_object_name is None else "%s_%s" % (object_name, center_object_name)) if dd.has_key(name): normalized_distance, i_center, j_center, good_mask = dd[name] else: d_to_edge = distance_to_edge(labels) if center_object_name is not None: # # Use the center of the centering objects to assign a center # to each labeled pixel using propagation # center_objects = workspace.object_set.get_objects(center_object_name) center_labels, cmask = cpo.size_similarly( labels, center_objects.segmented) pixel_counts = fix(scind.sum( np.ones(center_labels.shape), center_labels, np.arange(1, np.max(center_labels) + 1, dtype=np.int32))) good = pixel_counts > 0 i, j = (centers_of_labels(center_labels) + .5).astype(int) ig = i[good] jg = j[good] lg = np.arange(1, len(i) + 1)[good] if center_choice == C_CENTERS_OF_OTHER: # # Reduce the propagation labels to the centers of # the centering objects # center_labels = np.zeros(center_labels.shape, int) center_labels[ig, jg] = lg cl, d_from_center = propagate(np.zeros(center_labels.shape), center_labels, labels != 0, 1) # # Erase the centers that fall outside of labels # cl[labels == 0] = 0 # # If objects are hollow or crescent-shaped, there may be # objects without center labels. As a backup, find the # center that is the closest to the center of mass. # missing_mask = (labels != 0) & (cl == 0) missing_labels = np.unique(labels[missing_mask]) if len(missing_labels): all_centers = centers_of_labels(labels) missing_i_centers, missing_j_centers = \ all_centers[:, missing_labels - 1] di = missing_i_centers[:, np.newaxis] - ig[np.newaxis, :] dj = missing_j_centers[:, np.newaxis] - jg[np.newaxis, :] missing_best = lg[np.argsort((di * di + dj * dj,))[:, 0]] best = np.zeros(np.max(labels) + 1, int) best[missing_labels] = missing_best cl[missing_mask] = best[labels[missing_mask]] # # Now compute the crow-flies distance to the centers # of these pixels from whatever center was assigned to # the object. # iii, jjj = np.mgrid[0:labels.shape[0], 0:labels.shape[1]] di = iii[missing_mask] - i[cl[missing_mask] - 1] dj = jjj[missing_mask] - j[cl[missing_mask] - 1] d_from_center[missing_mask] = np.sqrt(di * di + dj * dj) else: # Find the point in each object farthest away from the edge. # This does better than the centroid: # * The center is within the object # * The center tends to be an interesting point, like the # center of the nucleus or the center of one or the other # of two touching cells. # i, j = maximum_position_of_labels(d_to_edge, labels, objects.indices) center_labels = np.zeros(labels.shape, int) center_labels[i, j] = labels[i, j] # # Use the coloring trick here to process touching objects # in separate operations # colors = color_labels(labels) ncolors = np.max(colors) d_from_center = np.zeros(labels.shape) cl = np.zeros(labels.shape, int) for color in range(1, ncolors + 1): mask = colors == color l, d = propagate(np.zeros(center_labels.shape), center_labels, mask, 1) d_from_center[mask] = d[mask] cl[mask] = l[mask] good_mask = cl > 0 if center_choice == C_EDGES_OF_OTHER: # Exclude pixels within the centering objects # when performing calculations from the centers good_mask = good_mask & (center_labels == 0) i_center = np.zeros(cl.shape) i_center[good_mask] = i[cl[good_mask] - 1] j_center = np.zeros(cl.shape) j_center[good_mask] = j[cl[good_mask] - 1] normalized_distance = np.zeros(labels.shape) if wants_scaled: total_distance = d_from_center + d_to_edge normalized_distance[good_mask] = (d_from_center[good_mask] / (total_distance[good_mask] + .001)) else: normalized_distance[good_mask] = \ d_from_center[good_mask] / maximum_radius dd[name] = [normalized_distance, i_center, j_center, good_mask] ngood_pixels = np.sum(good_mask) good_labels = labels[good_mask] bin_indexes = (normalized_distance * bin_count).astype(int) bin_indexes[bin_indexes > bin_count] = bin_count labels_and_bins = (good_labels - 1, bin_indexes[good_mask]) histogram = coo_matrix((pixel_data[good_mask], labels_and_bins), (nobjects, bin_count + 1)).toarray() sum_by_object = np.sum(histogram, 1) sum_by_object_per_bin = np.dstack([sum_by_object] * (bin_count + 1))[0] fraction_at_distance = histogram / sum_by_object_per_bin number_at_distance = coo_matrix((np.ones(ngood_pixels), labels_and_bins), (nobjects, bin_count + 1)).toarray() object_mask = number_at_distance > 0 sum_by_object = np.sum(number_at_distance, 1) sum_by_object_per_bin = np.dstack([sum_by_object] * (bin_count + 1))[0] fraction_at_bin = number_at_distance / sum_by_object_per_bin mean_pixel_fraction = fraction_at_distance / (fraction_at_bin + np.finfo(float).eps) masked_fraction_at_distance = masked_array(fraction_at_distance, ~object_mask) masked_mean_pixel_fraction = masked_array(mean_pixel_fraction, ~object_mask) # Anisotropy calculation. Split each cell into eight wedges, then # compute coefficient of variation of the wedges' mean intensities # in each ring. # # Compute each pixel's delta from the center object's centroid i, j = np.mgrid[0:labels.shape[0], 0:labels.shape[1]] imask = i[good_mask] > i_center[good_mask] jmask = j[good_mask] > j_center[good_mask] absmask = (abs(i[good_mask] - i_center[good_mask]) > abs(j[good_mask] - j_center[good_mask])) radial_index = (imask.astype(int) + jmask.astype(int) * 2 + absmask.astype(int) * 4) statistics = [] for bin in range(bin_count + (0 if wants_scaled else 1)): bin_mask = (good_mask & (bin_indexes == bin)) bin_pixels = np.sum(bin_mask) bin_labels = labels[bin_mask] bin_radial_index = radial_index[bin_indexes[good_mask] == bin] labels_and_radii = (bin_labels - 1, bin_radial_index) radial_values = coo_matrix((pixel_data[bin_mask], labels_and_radii), (nobjects, 8)).toarray() pixel_count = coo_matrix((np.ones(bin_pixels), labels_and_radii), (nobjects, 8)).toarray() mask = pixel_count == 0 radial_means = masked_array(radial_values / pixel_count, mask) radial_cv = np.std(radial_means, 1) / np.mean(radial_means, 1) radial_cv[np.sum(~mask, 1) == 0] = 0 for measurement, feature, overflow_feature in ( (fraction_at_distance[:, bin], MF_FRAC_AT_D, OF_FRAC_AT_D), (mean_pixel_fraction[:, bin], MF_MEAN_FRAC, OF_MEAN_FRAC), (np.array(radial_cv), MF_RADIAL_CV, OF_RADIAL_CV)): if bin == bin_count: measurement_name = overflow_feature % image_name else: measurement_name = feature % (image_name, bin + 1, bin_count) measurements.add_measurement(object_name, measurement_name, measurement) if feature in heatmaps: heatmaps[feature][bin_mask] = measurement[bin_labels - 1] radial_cv.mask = np.sum(~mask, 1) == 0 bin_name = str(bin + 1) if bin < bin_count else "Overflow" statistics += [(image_name, object_name, bin_name, str(bin_count), round(np.mean(masked_fraction_at_distance[:, bin]), 4), round(np.mean(masked_mean_pixel_fraction[:, bin]), 4), round(np.mean(radial_cv), 4))] return statistics
def run(self, workspace): # # Get some things we need from the workspace # measurements = workspace.measurements object_set = workspace.object_set # # Get the objects # objects_name = self.objects_name.value objects = object_set.get_objects(objects_name) # # First, I do it the (1) way to show how that code should look. # Later, I do it the (3) way and that will work even if objects.has_ijv # is False. if self.method == SUPPORT_BASIC: labels = objects.segmented # # The indices are the integer values representing each of the objects # in the labels matrix. scipy.ndimage functions often take an optional # argument that tells them which objects should be analyzed. # For instance, scipy.ndimage.mean takes an input image, a labels matrix # and the indices. If you don't supply the indices, it will just take # the mean of all labeled pixels, returning a single number. # indices = objects.indices # # Find the labeled pixels using labels != 0 # foreground = labels != 0 # # use scipy.ndimage.distance_transform_edt to find the distance of # every foreground pixel from the object edge # distance = scipy.ndimage.distance_transform_edt(foreground) # # call scipy.ndimage.mean(distance, labels, indices) to find the # mean distance in each object from its edge # values = scipy.ndimage.mean(distance, labels, indices) # # record the measurement using measurements.add_measurement # with an object name of "objects_name" and a measurement name # of M_MEAN_DISTANCE # measurements.add_measurement(objects_name, M_MEAN_DISTANCE, values) elif self.method == SUPPORT_OVERLAPPING: # # I'll use objects.get_labels to get labels matrices. This involves # a little extra work coallating the values, but not so bad. # # First of all, labels indices start at 1, but arrays start at # zero, so for "values", I'm going to cheat and waste values[0]. # Later, I'll only use values[1:] # values = np.zeros(objects.count + 1) # # Now for the loop # for labels, indices in objects.get_labels(): foreground = labels != 0 distance = scipy.ndimage.distance_transform_edt(foreground) v1 = scipy.ndimage.mean(distance, labels, indices) # # We copy the values above into the appropriate slots # values[indices] = v1 measurements.add_measurement(objects_name, M_MEAN_DISTANCE, values[1:]) else: # # It's just a little expensive finding out which labels are # touching others. The trick here is to use a function from # cpmorphology called "color_labels". This is akin to the # four color theorem - you want to color objects so that no # two adjacent ones have the same color. # # After we've done that, we process each of the colors in turn, # knowing that each object is colored only once and none of its # neighbors have the same color. # # This is a good demo of why Python and Numpy are good choices # for image processing. We're handling some pretty abstract # concepts in just a few lines of code and the result, I hope, # is clear and readable. # from centrosome.cpmorphology import color_labels values = np.zeros(objects.count + 1) for labels, indices in objects.get_labels(): clabels = color_labels(labels) # # np.unique returns the unique #s in an array. # colors = np.unique(clabels) for color in colors: # 0 = background, so ignore it. if color == 0: continue # # Ok, here's a trick. clabels == color gets converted # to either 1 (is the current color) or 0 (is not) and # we can use that to mask only the labels for the current # color by multiplying (0 * anything = 0) # foreground = clabels == color mini_labels = labels * foreground distance = scipy.ndimage.distance_transform_edt(foreground) # # And here's another trick - scipy.ndimage.mean returns # NaN for any index that doesn't appear because the # mean isn't computable. How lucky! # v1 = scipy.ndimage.mean(distance, mini_labels, indices) good_v1 = ~np.isnan(v1) values[indices[good_v1]] = v1[good_v1] measurements.add_measurement(objects_name, M_MEAN_DISTANCE, values[1:])