-
-import pandas as pd
-import numpy as np
-import libtiff
-import skimage.exposure
-import skimage.feature
-import skimage.filters
-import skimage.morphology
-
-def make_mask(zstack, nbins=32768):
- thresh = skimage.filters.threshold_otsu(zstack, nbins=32768)
- return zstack > thresh
-
-def build_blob_array(stack):
- data = [[[False for i in range(len(stack[0][0]))] for j in range(len(stack[0]))] for k in range(len(stack))]
- for im in range(len(stack))): #was stack
- print "processing " + str(im)
- yx = skimage.exposure.equalize_adapthist(stack[im], clip_limit=0.00025, nbins=32768)
-
- zmx = np.max(yx)
- zmn = np.min(yx)
- yx = (yx - zmn) / (zmx - zmn)
-
- blobs_log = skimage.feature.blob_log(yx, min_sigma=1, max_sigma=3, num_sigma=20, threshold=0.025)
- print str(len(blobs_log)) + " blobs found"
- if len(blobs_log) > 1:
- blobs_log[:, 2] = blobs_log[:, 2] * math.sqrt(2)
- for i in blobs_log:
- data[im][int(i[0])][int(i[1])] = True
- return data
-
-def make_watershed(mask):
- distance = ndi.distance_transform_edt(mask)
- local_max = skimage.feature.peak_local_max(distance, indices=False, footprint=np.ones((31, 31)), labels=mask)
- markers = ndi.label(local_max)[0]
- labels = skimage.morphology.watershed(-distance, markers, mask=mask)
- return labels
-
-def make_gene_layers(gene_tif):
- gene_z = gene_tif.get_tiff_array()
- gene_s = np.arange(0, len(gene_z)+1, len(gene_z) / 6)
- gene_r = zip(gene_s[:-1], (gene_s + 1)[1:])
- gene_y = np.array(gene_z[gene_r[2][0]:gene_r[2][1]], dtype=np.uint16)
- return gene_y
-
-def make_nissl_watershed(nissl_tif):
- nissl_z = nissl_tif.get_tiff_array()
- nissl_s = np.arange(0, len(nissl_z)+1, len(nissl_z) / 6)
- nissl_r = zip(nissl_s[:-1], (nissl_s + 1)[1:])
- nissl_y = np.array(nissl_z[nissl_r[1][0]:nissl_r[1][1]], dtype=np.uint16)
- nissl_watershed = [[] for k in range(len(nissl_y))]
-
- for i in range(len(nissl_y)):
- mask = make_mask(nissl_y[i])
- watershed = make_watershed(mask)
- nissl_watershed[i] = watershed
- return nissl_watershed
-
-def correct_labels(nissl_watershed):
- nissl_watershed_corrected = [[[0 for i in range(len(nissl_watershed[0][0]))] for j in range(len(nissl_watershed[0]))] for k in range(len(nissl_watershed))]
- nissl_watershed_corrected[0] = [i for i in nissl_watershed[0]]
- nissl_watershed_corrected = np.asarray(nissl_watershed_corrected)
-
- for k in range(1, len(nissl_watershed_corrected)):
- cell_number_max = nissl_watershed_corrected[k-1].max() #changed to corrected to keep accurate count
- cell_number_two_max = nissl_watershed[k].max()
- and_map = np.logical_and(nissl_watershed[k-1], nissl_watershed[k])
- and_map_pos = np.where(and_map)
- cell_set = set(nissl_watershed[k-1][np.where(and_map)])
- inter_layer_dict = dict()
- for i in range(len(and_map_pos[0])):
- inter_layer_dict[nissl_watershed[k][and_map_pos[0][i]][and_map_pos[1][i]]] = nissl_watershed[k-1][and_map_pos[0][i]][and_map_pos[1][i]]
- counter = 0
- inter_layer_dict[0] = 0
- for i in range(1, cell_number_two_max + 1):
- if not i in inter_layer_dict:
- counter += 1
- inter_layer_dict[i] = cell_number_max + counter
-
-
- for j in range(1024):
- for i in range(1024):
- nissl_watershed_corrected[k][i][j] = inter_layer_dict[nissl_watershed[k][i][j]]
-
- return nissl_watershed_corrected