def segment_glomeruli2d(input_file, tissue_mask_file, output_file, voxel_xy):
kmask = io.imread(tissue_mask_file)
if kmask.max() == 0:
tifffile.imsave(output_file, kmask, compress=5)
return
# normalize image
img = io.imread(input_file)
img = ndimage.median_filter(img, 3)
img = img * 255. / img.max()
# remove all intensity variations larger than maximum radius of a glomerulus
d = mahotas.disk(int(float(glomeruli_maxrad) / voxel_xy))
img = img - mahotas.open(img.astype(np.uint8), d)
img = img * 255. / img.max()
ch = img[np.where(kmask > 0)]
# segment glomeruli by otsu thresholding only if this threshold is higher than the 75-th percentile in the kidney mask
t = mahotas.otsu(img.astype(np.uint8))
cells = None
if t > np.percentile(ch, 75) * 1.5:
cells = img > t
cells[np.where(kmask == 0)] = 0
cells = mahotas.open(
cells, mahotas.disk(int(float(glomeruli_minrad) / 2. / voxel_xy)))
else:
cells = np.zeros_like(img)
tifffile.imsave(output_file, img_as_ubyte(cells), compress=5)
def generate_correct(image, prob, segmentation):
hist = Util.get_histogram(segmentation.astype(np.uint64))
labels = range(1, len(hist)) # do not include zeros
np.random.shuffle(labels)
for l in labels:
binary_mask = Util.threshold(segmentation, l)
borders = mh.labeled.borders(binary_mask, Bc=mh.disk(2))
labeled_borders = skimage.measure.label(borders)
labeled_borders[borders == 0] = 0
relabeled_borders, no_relabeled_borders = mh.labeled.relabel(
labeled_borders.astype(np.uint16))
for border in range(1, no_relabeled_borders + 1):
isolated_border = Util.threshold(relabeled_borders, border)
patches = Patch.analyze_border(image, prob, binary_mask,
binary_mask.invert(),
isolated_border)
for s in patches:
yield s
yield Patch.fliplr(s)
yield Patch.flipud(s)
yield Patch.rotate(s, 90)
yield Patch.rotate(s, 180)
yield Patch.rotate(s, 270)
def extract(self):
'''Extracts neighbour features.
Returns
-------
pandas.DataFrame
extracted feature values for each object in `label_image`
'''
# Create an empty dataset in case no objects were detected
logger.info('extract neighbour features')
features = list()
for obj in self.object_ids:
pad = max(self.neighbour_distance, self.touching_distance)
object_image = self._get_bbox_containing_neighbours(obj, pad)
# dilate the current object
object_image_dilate = mh.dilate(object_image == obj,
Bc=mh.disk(
self.neighbour_distance))
# mask the corresponding region of the label image
object_image_mask = np.copy(object_image)
object_image_mask[object_image_dilate == 0] = 0
object_image_mask[object_image == obj] = 0
neighbour_ids = np.unique(object_image_mask)
unique_values = neighbour_ids[np.nonzero(neighbour_ids)].tolist()
neighbour_count = len(unique_values)
# save these unique values as a string
if neighbour_count == 0:
neighbour_string = '.'
else:
neighbour_string = '.'.join(str(x) for x in unique_values)
# create an inverted image of the surrounding cells
neighbours = np.zeros_like(object_image)
for n in unique_values:
neighbours += mh.dilate(object_image == n)
# calculate the distance from each pixel of object to neighbours
dist = ndi.morphology.distance_transform_edt(
np.invert(neighbours > 0))
# select perimeter pixels whose distance to neighbours is
# less than threshold touching distance
perimeter_image = mh.bwperim(object_image == obj)
dist[perimeter_image == 0] = 0
dist[dist > self.touching_distance] = 0
fraction_touching = np.count_nonzero(dist) / float(
np.count_nonzero(perimeter_image))
values = [neighbour_count, neighbour_string, fraction_touching]
features.append(values)
return pd.DataFrame(features,
columns=self.names,
index=self.object_ids)
def segment_layer(filename, params): ''' Segment one layer in a stack ''' #extract pixel size in xy and z xsize, zsize = extract_zoom(params.folder) #load image img = tifffile.imread(params.inputfolder + params.folder + filename) #normalize image img = ndimage.median_filter(img, 3) per_low = np.percentile(img, 5) img[img < per_low] = per_low img = img - img.min() per_high = np.percentile(img, 99) img[img > per_high] = per_high img = img*255./img.max() imgf = ndimage.gaussian_filter(img*1., 30./xsize).astype(np.uint8) kmask = (imgf > mahotas.otsu(imgf.astype(np.uint8)))*255. sizefactor = 10 small = ndimage.interpolation.zoom(kmask, 1./sizefactor) #scale the image to a smaller size rad = int(300./xsize) small_ext = np.zeros([small.shape[0] + 4*rad, small.shape[1] + 4*rad]) small_ext[2*rad : 2*rad + small.shape[0], 2*rad : 2*rad + small.shape[1]] = small small_ext = mahotas.close(small_ext.astype(np.uint8), mahotas.disk(rad)) small = small_ext[2*rad : 2*rad + small.shape[0], 2*rad : 2*rad + small.shape[1]] small = mahotas.close_holes(small)*1. small = small*255./small.max() kmask = ndimage.interpolation.zoom(small, sizefactor) #scale back to normal size kmask = normalize(kmask) kmask = (kmask > mahotas.otsu(kmask.astype(np.uint8)))*255. #remove artifacts of interpolation if np.median(imgf[np.where(kmask > 0)]) < (np.median(imgf[np.where(kmask == 0)]) + 1)*3: kmask = np.zeros_like(kmask) #save indices of the kidney mask # ind = np.where(kmask > 0) # ind = np.array(ind) # np.save(params.inputfolder + '../segmented/masks/' + params.folder + filename[:-4] + '.npy', ind) #save outlines im = np.zeros([img.shape[0], img.shape[1], 3]) img = tifffile.imread(params.inputfolder + params.folder + filename) im[:,:,0] = im[:,:,1] = im[:,:,2] = np.array(img) output = overlay(kmask, im, (255,0,0), borders = True) tifffile.imsave(params.inputfolder + '../segmented/outlines/' + params.folder + filename[:-4] + '.tif', (output).astype(np.uint8))
def test_eccentricity():
D = mh.disk(32, 2)
ecc = mahotas.features.shape.eccentricity(D)
assert 0 <= ecc < .01
Index = np.indices((33,33)).astype(float)
Index -= 15
X,Y = Index
ellipse = ((X**2+2*Y**2) < 12**2)
assert 0 < mahotas.features.shape.eccentricity(ellipse) < 1
def test_eccentricity():
D = mh.disk(32, 2)
ecc = mahotas.features.shape.eccentricity(D)
assert 0 <= ecc < .01
Index = np.indices((33, 33)).astype(float)
Index -= 15
X, Y = Index
ellipse = ((X**2 + 2 * Y**2) < 12**2)
assert 0 < mahotas.features.shape.eccentricity(ellipse) < 1
def segment_tissue2d(input_file, output_file, voxel_xy):
# load image
img = io.imread(input_file)
# normalize image
img = ndimage.median_filter(img, 3)
img = img * 255. / img.max()
##segment kidney tissue
sizefactor = 10.
small = ndimage.interpolation.zoom(
img, 1. / sizefactor) # scale the image to a smaller size
imgf = ndimage.gaussian_filter(small, 3. / voxel_xy) # Gaussian filter
median = np.percentile(imgf, 40) # 40-th percentile for thresholding
kmask = imgf > median * 1.5 # thresholding
kmask = mahotas.dilate(kmask, mahotas.disk(5))
kmask = mahotas.close_holes(kmask) # closing holes
kmask = mahotas.erode(kmask, mahotas.disk(5)) * 255
# remove objects that are darker than 2*percentile
l, n = ndimage.label(kmask)
llist = np.unique(l)
if len(llist) > 2:
means = ndimage.mean(imgf, l, llist)
bv = llist[np.where(means < median * 2)]
ix = np.in1d(l.ravel(), bv).reshape(l.shape)
kmask[ix] = 0
kmask = ndimage.interpolation.zoom(kmask,
sizefactor) # scale back to normal size
kmask = normalize(kmask)
kmask = (kmask > mahotas.otsu(kmask.astype(np.uint8))
) # remove artifacts of interpolation
tifffile.imsave(output_file, img_as_ubyte(kmask), compress=5)
def random_watershed(array, speed_image, border_seeds=False, erode=False):
'''
'''
copy_array = np.array(array, dtype=np.bool)
if erode:
for i in range(10):
copy_array = mh.erode(copy_array)
seed_array = np.array(copy_array)
if border_seeds:
seed_array = mh.labeled.border(copy_array, 1, 0, Bc=mh.disk(7))
coords = zip(*np.where(seed_array==1))
if len(coords) == 0:
# print 'err'
return np.zeros(array.shape)
seed1_ = None
seed2_ = None
max_distance = -np.inf
for i in range(10):
seed1 = random.choice(coords)
seed2 = random.choice(coords)
d = distance.euclidean(seed1, seed2)
if max_distance < d:
max_distance = d
seed1_ = seed1
seed2_ = seed2
seeds = np.zeros(array.shape, dtype=np.uint8)
seeds[seed1_[0], seed1_[1]] = 1
seeds[seed2_[0], seed2_[1]] = 2
for i in range(8):
seeds = mh.dilate(seeds)
# Util.view(seeds,large=True)
# print speed_image.shape, seeds.shape
ws = mh.cwatershed(speed_image, seeds)
ws[array == 0] = 0
return ws
def mahotas_clean_up_seg(input_jacobian, frame_num):
import mahotas as mh
dsk = mh.disk(7)
thresh_r = 0.1
thresh_g = 1
size_cutoff = 200
thresholded_jacobian = (np.int32(np.log(1+input_jacobian[frame_num][0])>thresh_r)+\
np.int32(np.log(1+input_jacobian[frame_num][1])>thresh_g))>0
thresholded_jacobian = mh.close_holes(thresholded_jacobian)
thresholded_jacobian = mh.erode(thresholded_jacobian, dsk)
labeled = mh.label(thresholded_jacobian)[0]
sizes = mh.labeled.labeled_size(labeled)
too_small = np.where(sizes < size_cutoff)
labeled = mh.labeled.remove_regions(labeled, too_small)
thresholded_jacobian = labeled > 0
thresholded_jacobian = mh.dilate(thresholded_jacobian, dsk)
return thresholded_jacobian
def show_segmentation_boundaries(input_image,seg_mask,frame_num,sizeX,sizeY):
import mahotas as mh
cmap = plt.get_cmap('gray')
#outlines
dsk = mh.disk(7)
boundaries = mh.dilate(seg_mask,dsk)-seg_mask
boundaries=boundaries>0
cmap_2_use = truncate_colormap(cmap,0.9,1)
border2overlay = np.ma.masked_where(boundaries ==0, boundaries)
x =mh.as_rgb(input_image[frame_num][0,:,:],input_image[frame_num][1,:,:],np.zeros((sizeY,sizeX)))
x[:,:,1][x[:,:,1]>240]=240
x[:,:,0][x[:,:,0]>240]=240
slice2show = [slice(0,sizeY),slice(0,sizeX)]
plt.imshow(x[slice2show]/np.array([240.,240.,1]).reshape(1,1,3))
plt.imshow(border2overlay[slice2show],cmap_2_use,alpha=0.8),plt.axis('off')
def mahotas_clean_up_seg(input_jacobian,frame_num):
import mahotas as mh
dsk = mh.disk(7)
thresh_r = 0.1
thresh_g = 1
size_cutoff = 200
thresholded_jacobian = (np.int32(np.log(1+input_jacobian[frame_num][0])>thresh_r)+\
np.int32(np.log(1+input_jacobian[frame_num][1])>thresh_g))>0
thresholded_jacobian = mh.close_holes(thresholded_jacobian)
thresholded_jacobian = mh.erode(thresholded_jacobian,dsk)
labeled = mh.label(thresholded_jacobian)[0]
sizes = mh.labeled.labeled_size(labeled)
too_small = np.where(sizes < size_cutoff)
labeled = mh.labeled.remove_regions(labeled, too_small)
thresholded_jacobian = labeled>0
thresholded_jacobian = mh.dilate(thresholded_jacobian,dsk)
return thresholded_jacobian
def generate_correct(image, prob, label):
'''
'''
labels = np.arange(len(Util.get_histogram(label)))[1:] # no zeros
np.random.shuffle(labels)
for l in labels:
# neighbors = PG.grab_neighbors(label, l)
# for n in neighbors:
# patches = PG.analyze_border(image, prob, label, l, n, sample_rate=10)
# for s in patches:
# yield s
# yield PG.fliplr(s)
# yield PG.flipud(s)
# yield PG.rotate(s, 90)
# yield PG.rotate(s, 180)
# yield PG.rotate(s, 270)
binary_mask = Util.threshold(label, l)
borders = mh.labeled.borders(binary_mask,Bc=mh.disk(2))
labeled_borders = skimage.measure.label(borders)
labeled_borders[borders==0] = 0
relabeled_borders, no_relabeled_borders = mh.labeled.relabel(labeled_borders.astype(np.uint16))
for border in range(1,no_relabeled_borders+1):
isolated_border = Util.threshold(relabeled_borders, border)
patches = PGNew.analyze_border(image, prob, binary_mask, isolated_border)
for s in patches:
yield s
yield PGNew.fliplr(s)
yield PGNew.flipud(s)
yield PGNew.rotate(s, 90)
yield PGNew.rotate(s, 180)
yield PGNew.rotate(s, 270)
def show_segmentation_boundaries(input_image, seg_mask, frame_num, sizeX,
sizeY):
import mahotas as mh
cmap = plt.get_cmap('gray')
#outlines
dsk = mh.disk(7)
boundaries = mh.dilate(seg_mask, dsk) - seg_mask
boundaries = boundaries > 0
cmap_2_use = truncate_colormap(cmap, 0.9, 1)
border2overlay = np.ma.masked_where(boundaries == 0, boundaries)
x = mh.as_rgb(input_image[frame_num][0, :, :],
input_image[frame_num][1, :, :], np.zeros((sizeY, sizeX)))
x[:, :, 1][x[:, :, 1] > 240] = 240
x[:, :, 0][x[:, :, 0] > 240] = 240
slice2show = [slice(0, sizeY), slice(0, sizeX)]
plt.imshow(x[slice2show] / np.array([240., 240., 1]).reshape(1, 1, 3))
plt.imshow(border2overlay[slice2show], cmap_2_use,
alpha=0.8), plt.axis('off')
def fix_single_merge(cnn, cropped_image, cropped_prob, cropped_binary, N=10, invert=True, dilate=True,
border_seeds=True, erode=False, debug=False, before_merge_error=None,
real_border=np.zeros((1,1)), oversampling=False, crop=True):
'''
invert: True/False for invert or gradient image
'''
bbox = mh.bbox(cropped_binary)
orig_cropped_image = np.array(cropped_image)
orig_cropped_prob = np.array(cropped_prob)
orig_cropped_binary = np.array(cropped_binary)
speed_image = None
if invert:
speed_image = Legacy.invert(cropped_image, smooth=True, sigma=2.5)
else:
speed_image = Legacy.gradient(cropped_image)
Util.view(speed_image, large=False, color=False)
dilated_binary = np.array(cropped_binary, dtype=np.bool)
if dilate:
for i in range(20):
dilated_binary = mh.dilate(dilated_binary)
Util.view(dilated_binary, large=False, color=False)
borders = np.zeros(cropped_binary.shape)
best_border_prediction = np.inf
best_border_image = np.zeros(cropped_binary.shape)
original_border = mh.labeled.border(cropped_binary, 1, 0, Bc=mh.disk(3))
results_no_border = []
predictions = []
borders = []
results = []
for n in range(N):
ws = Legacy.random_watershed(dilated_binary, speed_image, border_seeds=border_seeds, erode=erode)
if ws.max() == 0:
continue
ws_label1 = ws.max()
ws_label2 = ws.max()-1
border = mh.labeled.border(ws, ws_label1, ws_label2)
# Util.view(ws, large=True)
# Util.view(border, large=True)
# print i, len(border[border==True])
#
# remove parts of the border which overlap with the original border
#
ws[cropped_binary == 0] = 0
# Util.view(ws, large=False, color=False)
ws_label1_array = Util.threshold(ws, ws_label1)
ws_label2_array = Util.threshold(ws, ws_label2)
eroded_ws1 = np.array(ws_label1_array, dtype=np.bool)
eroded_ws2 = np.array(ws_label2_array, dtype=np.bool)
if erode:
for i in range(5):
eroded_ws1 = mh.erode(eroded_ws1)
# Util.view(eroded_ws, large=True, color=False)
dilated_ws1 = np.array(eroded_ws1)
for i in range(5):
dilated_ws1 = mh.dilate(dilated_ws1)
for i in range(5):
eroded_ws2 = mh.erode(eroded_ws2)
# Util.view(eroded_ws, large=True, color=False)
dilated_ws2 = np.array(eroded_ws2)
for i in range(5):
dilated_ws2 = mh.dilate(dilated_ws2)
new_ws = np.zeros(ws.shape, dtype=np.uint8)
new_ws[dilated_ws1 == 1] = ws_label1
new_ws[dilated_ws2 == 1] = ws_label2
ws = new_ws
# Util.view(new_ws, large=True, color=True)
# ws[original_border == 1] = 0
prediction = Patch.grab_group_test_and_unify(cnn, cropped_image, cropped_prob, ws, ws_label1, ws_label2, oversampling=oversampling)
if prediction == -1 or prediction >= .5:
# invalid
continue
# here we have for one border
# the border
# the prediction
# borders.append(border)
# predictions.append(prediction)
results.append((prediction, border))
return results
def test_perimeter():
for r in (40, 80, 160):
disk = mh.disk(r, 2)
p = mahotas.labeled.perimeter(disk)
p_exact = r * np.pi * 2
assert .9 < (p / p_exact) < 1.1
def test_perimeter():
for r in (40,80,160):
disk = mh.disk(r, 2)
p = mahotas.labeled.perimeter(disk)
p_exact = r*np.pi*2
assert .9 < (p/p_exact) < 1.1
def segment_layer(filename, params):
'''
Segment one layer in a stack
'''
start = time.time()
#extract pixel size in xy and z
xsize, zsize = extract_zoom(params.folder)
#load image
img = tifffile.imread(params.inputfolder + params.folder + filename)
#normalize image
img = ndimage.median_filter(img, 3)
img = img * 255. / img.max()
##segment kidney tissue
sizefactor = 10.
small = ndimage.interpolation.zoom(
img, 1. / sizefactor) #scale the image to a smaller size
imgf = ndimage.gaussian_filter(small, 3. / xsize) #Gaussian filter
median = np.percentile(imgf, 40) #40-th percentile for thresholding
kmask = imgf > median * 1.5 #thresholding
kmask = mahotas.dilate(kmask, mahotas.disk(5))
kmask = mahotas.close_holes(kmask) #closing holes
kmask = mahotas.erode(kmask, mahotas.disk(5)) * 255
#remove objects that are darker than 2*percentile
l, n = ndimage.label(kmask)
llist = np.unique(l)
if len(llist) > 2:
means = ndimage.mean(imgf, l, llist)
bv = llist[np.where(means < median * 2)]
ix = np.in1d(l.ravel(), bv).reshape(l.shape)
kmask[ix] = 0
kmask = ndimage.interpolation.zoom(kmask,
sizefactor) #scale back to normal size
kmask = normalize(kmask)
kmask = (kmask > mahotas.otsu(kmask.astype(
np.uint8))) * 255. #remove artifacts of interpolation
#save indices of the kidney mask
ind = np.where(kmask > 0)
ind = np.array(ind)
np.save(
params.inputfolder + '../segmented/masks/kidney/' + params.folder +
filename[:-4] + '.npy', ind)
#segment glomeruli, if there is a kidney tissue
if kmask.max() > 0:
#remove all intensity variations larger than maximum radius of a glomerulus
d = mahotas.disk(int(float(params.maxrad) / xsize))
img = img - mahotas.open(img.astype(np.uint8), d)
img = img * 255. / img.max()
ch = img[np.where(kmask > 0)]
#segment glomeruli by otsu thresholding only if this threshold is higher than the 75-th percentile in the kidney mask
t = mahotas.otsu(img.astype(np.uint8))
if t > np.percentile(ch, 75) * 1.5:
cells = img > t
cells[np.where(kmask == 0)] = 0
cells = mahotas.open(
cells, mahotas.disk(int(float(params.minrad) / 2. / xsize)))
else:
cells = np.zeros_like(img)
else:
cells = np.zeros_like(img)
#save indices of the glomeruli mask
ind = np.where(cells > 0)
ind = np.array(ind)
np.save(
params.inputfolder + '../segmented/masks/glomeruli/' + params.folder +
filename[:-4] + '.npy', ind)
def fix_single_merge(cnn,
cropped_image,
cropped_prob,
cropped_binary,
N=10,
invert=True,
dilate=True,
border_seeds=True,
erode=False,
debug=False,
before_merge_error=None,
real_border=np.zeros((1, 1)),
oversampling=False,
crop=True):
'''
invert: True/False for invert or gradient image
'''
bbox = mh.bbox(cropped_binary)
orig_cropped_image = np.array(cropped_image)
orig_cropped_prob = np.array(cropped_prob)
orig_cropped_binary = np.array(cropped_binary)
speed_image = None
if invert:
speed_image = Util.invert(cropped_image, smooth=True, sigma=2.5)
else:
speed_image = Util.gradient(cropped_image)
dilated_binary = np.array(cropped_binary, dtype=np.bool)
if dilate:
for i in range(20):
dilated_binary = mh.dilate(dilated_binary)
# Util.view(dilated_binary, large=True)
borders = np.zeros(cropped_binary.shape)
best_border_prediction = np.inf
best_border_image = np.zeros(cropped_binary.shape)
original_border = mh.labeled.border(cropped_binary,
1,
0,
Bc=mh.disk(3))
results_no_border = []
predictions = []
for n in range(N):
ws = Util.random_watershed(dilated_binary,
speed_image,
border_seeds=border_seeds,
erode=erode)
if ws.max() == 0:
continue
ws_label1 = ws.max()
ws_label2 = ws.max() - 1
border = mh.labeled.border(ws, ws_label1, ws_label2)
# Util.view(ws, large=True)
# Util.view(border, large=True)
# print i, len(border[border==True])
#
# remove parts of the border which overlap with the original border
#
ws[cropped_binary == 0] = 0
# Util.view(ws, large=False, color=False)
ws_label1_array = Util.threshold(ws, ws_label1)
ws_label2_array = Util.threshold(ws, ws_label2)
eroded_ws1 = np.array(ws_label1_array, dtype=np.bool)
eroded_ws2 = np.array(ws_label2_array, dtype=np.bool)
if erode:
for i in range(5):
eroded_ws1 = mh.erode(eroded_ws1)
# Util.view(eroded_ws, large=True, color=False)
dilated_ws1 = np.array(eroded_ws1)
for i in range(5):
dilated_ws1 = mh.dilate(dilated_ws1)
for i in range(5):
eroded_ws2 = mh.erode(eroded_ws2)
# Util.view(eroded_ws, large=True, color=False)
dilated_ws2 = np.array(eroded_ws2)
for i in range(5):
dilated_ws2 = mh.dilate(dilated_ws2)
new_ws = np.zeros(ws.shape, dtype=np.uint8)
new_ws[dilated_ws1 == 1] = ws_label1
new_ws[dilated_ws2 == 1] = ws_label2
ws = new_ws
# Util.view(new_ws, large=True, color=True)
# ws[original_border == 1] = 0
prediction = Patch.grab_group_test_and_unify(
cnn,
cropped_image,
cropped_prob,
ws,
ws_label1,
ws_label2,
oversampling=oversampling)
if prediction == -1:
# invalid
continue
# if (prediction < best_border_prediction):
# best_border_prediction = prediction
# best_border_image = border
# print 'new best', n, prediction
best_border_image = border
borders += (border * prediction)
result = np.array(cropped_binary)
best_border_image[result == 0] = 0
result[best_border_image == 1] = 2
result = skimage.measure.label(result)
result_no_border = np.array(result)
result_no_border[best_border_image == 1] = 0
predictions.append(prediction)
results_no_border.append(result_no_border)
# result = np.array(cropped_binary)
# best_border_image[result==0] = 0
# result[best_border_image==1] = 2
# result = skimage.measure.label(result)
# result_no_border = np.array(result)
# result_no_border[best_border_image==1] = 0
# result_no_border = mh.croptobbox(result_no_border)
# if before_merge_error == None:
# continue
# print result_no_border.shape, before_merge_error.shape
# if before_merge_error.shape[0] != result_no_border.shape[0] or before_merge_error.shape[1] != result_no_border.shape[1]:
# result_no_border = np.resize(result_no_border, before_merge_error.shape)
# print 'vi', Util.vi(before_merge_error.astype(np.uint8), result_no_border.astype(np.uint8))
# if debug:
# Util.view(ws, text=str(i) + ' ' + str(prediction))
result = np.array(cropped_binary)
best_border_image[result == 0] = 0
result[best_border_image == 1] = 2
result = skimage.measure.label(result)
result_no_border = np.array(result)
result_no_border[best_border_image == 1] = 0
return borders, best_border_image, result, result_no_border, results_no_border, predictions
def main(image,
mask,
threshold=150,
bead_size=2,
superpixel_size=4,
close_surface=False,
close_disc_size=8,
plot=False):
'''Converts an image stack with labelled cell surface to a cell
`volume` image
Parameters
----------
image: numpy.ndarray[Union[numpy.uint8, numpy.uint16]]
grayscale image in which beads should be detected (3D)
mask: numpy.ndarray[Union[numpy.int32, numpy.bool]]
binary or labeled image of cell segmentation (2D)
threshold: int, optional
intensity of bead (default: ``150``)
bead_size: int, optional
minimal size of bead (default: ``2``)
superpixel_size: int, optional
size of superpixels for searching the 3D position of a bead
close_surface: bool, optional
whether the interpolated surface should be morphologically closed
close_disc_size: int, optional
size in pixels of the disc used to morphologically close the
interpolated surface
plot: bool, optional
whether a plot should be generated (default: ``False``)
Returns
-------
jtmodules.generate_volume_image.Output
'''
n_slices = image.shape[-1]
logger.debug('input image has size %d in last dimension', n_slices)
logger.debug('mask beads inside cell')
beads_outside_cell = np.copy(image)
for iz in range(n_slices):
beads_outside_cell[mask > 0, iz] = 0
logger.debug('search for 3D position of beads outside cell')
slide = np.argmax(beads_outside_cell, axis=2)
slide[slide > np.percentile(slide[mask == 0], 20)] = 0
logger.debug('determine surface of slide')
slide_coordinates = array_to_coordinate_list(slide)
bottom_surface = fit_plane(
subsample_coordinate_list(slide_coordinates, 2000))
logger.debug('detect_beads in 2D')
mip = np.max(image, axis=-1)
try:
# TODO: use LOG filter???
beads, beads_centroids = detect_blobs(image=mip,
mask=np.invert(mask > 0),
threshold=threshold,
min_area=bead_size)
except:
logger.warn('detect_blobs failed, returning empty volume image')
volume_image = np.zeros(shape=mask.shape, dtype=image.dtype)
figure = str()
return Output(volume_image, figure)
n_beads = np.count_nonzero(beads_centroids)
logger.info('found %d beads on cells', n_beads)
if n_beads == 0:
logger.warn('empty volume image')
volume_image = np.zeros(shape=mask.shape, dtype=image.dtype)
else:
logger.debug('locate beads in 3D')
beads_coords_3D = locate_in_3D(image=image,
mask=beads_centroids,
bin_size=superpixel_size)
logger.info('interpolate cell surface')
volume_image = interpolate_surface(coords=beads_coords_3D,
output_shape=np.shape(image[:, :,
1]),
method='linear')
volume_image = volume_image.astype(image.dtype)
if (close_surface is True):
import mahotas as mh
logger.info('morphological closing of cell surface')
volume_image = mh.close(volume_image, Bc=mh.disk(close_disc_size))
volume_image[mask == 0] = 0
if plot:
logger.debug('convert bottom surface plane to image for plotting')
bottom_surface_image = np.zeros(slide.shape, dtype=np.uint8)
for ix in range(slide.shape[0]):
for iy in range(slide.shape[1]):
bottom_surface_image[ix, iy] = plane(ix, iy, bottom_surface.x)
logger.info('create plot')
from jtlib import plotting
plots = [
plotting.create_intensity_image_plot(mip, 'ul', clip=True),
plotting.create_intensity_image_plot(bottom_surface_image,
'll',
clip=True),
plotting.create_intensity_image_plot(volume_image, 'ur', clip=True)
]
figure = plotting.create_figure(plots,
title='Convert stack to volume image')
else:
figure = str()
return Output(volume_image, figure)
def split_new(image, binary):
'''
'''
bbox = mh.bbox(binary)
sub_image = np.array(image[bbox[0]:bbox[1], bbox[2]:bbox[3]])
sub_binary = np.array(binary[bbox[0]:bbox[1], bbox[2]:bbox[3]])
sub_binary_border = mh.labeled.borders(sub_binary, Bc=mh.disk(3))
sub_binary = mh.erode(sub_binary.astype(np.bool))
for e in range(5):
sub_binary = mh.erode(sub_binary)
# sub_binary = mh.erode(sub_binary)
if sub_image.shape[0] < 2 or sub_image.shape[1] < 2:
return np.zeros(binary.shape, dtype=np.bool), np.zeros(binary.shape, dtype=np.bool)
#
# smooth the image
#
sub_image = mh.gaussian_filter(sub_image, 3.5)
grad_x = np.gradient(sub_image)[0]
grad_y = np.gradient(sub_image)[1]
grad = np.add(np.abs(grad_x), np.abs(grad_y))
grad -= grad.min()
grad /= grad.max()
grad *= 255
grad = grad.astype(np.uint8)
coords = zip(*np.where(sub_binary==1))
if len(coords) < 2:
print 'STRAAAAANGE'
return np.zeros(binary.shape, dtype=np.bool), np.zeros(binary.shape, dtype=np.bool)
seed1 = random.choice(coords)
seed2 = random.choice(coords)
seeds = np.zeros(sub_binary.shape, dtype=np.uint64)
seeds[seed1] = 1
seeds[seed2] = 2
for i in range(10):
seeds = mh.dilate(seeds)
ws = mh.cwatershed(grad, seeds)
ws[sub_binary==0] = 0
# ws_relabeled = skimage.measure.label(ws.astype(np.uint8))
# ws_relabeled[sub_binary==0] = 0
# max_label = ws_relabeled.max()
# plt.figure()
# imshow(ws)
binary_mask = Util.threshold(ws, ws.max())
border = mh.labeled.border(ws, ws.max(), ws.max()-1, Bc=mh.disk(2))
# border[sub_binary_border == 1] = 0 # remove any "real" border pixels
# plt.figure()
# imshow(binary_mask)
# plt.figure()
# imshow(border)
large_label = np.zeros(binary.shape, dtype=np.bool)
large_border = np.zeros(binary.shape, dtype=np.bool)
large_label[bbox[0]:bbox[1], bbox[2]:bbox[3]] = binary_mask
large_border[bbox[0]:bbox[1], bbox[2]:bbox[3]] = border
return large_label, large_border
def split_label(image, binary):
bbox = mh.bbox(binary)
sub_image = np.array(image[bbox[0]:bbox[1], bbox[2]:bbox[3]])
sub_binary = np.array(binary[bbox[0]:bbox[1], bbox[2]:bbox[3]])
sub_binary_border = mh.labeled.borders(sub_binary, Bc=mh.disk(3))
sub_binary = mh.erode(sub_binary.astype(np.bool))
for e in range(15):
sub_binary = mh.erode(sub_binary)
# # sub_binary = mh.erode(sub_binary)
if sub_binary.shape[0] < 2 or sub_binary.shape[1] < 2:
return np.zeros(binary.shape,
dtype=np.bool), np.zeros(binary.shape,
dtype=np.bool)
#
# smooth the image
#
sub_image = mh.gaussian_filter(sub_image, 3.5)
grad_x = np.gradient(sub_image)[0]
grad_y = np.gradient(sub_image)[1]
grad = np.add(np.abs(grad_x), np.abs(grad_y))
grad -= grad.min()
grad /= grad.max()
grad *= 255
grad = grad.astype(np.uint8)
coords = zip(*np.where(sub_binary == 1))
if len(coords) < 2:
# print 'STRAAAAANGE'
return np.zeros(binary.shape,
dtype=np.bool), np.zeros(binary.shape,
dtype=np.bool)
seed1 = random.choice(coords)
seed2 = random.choice(coords)
seeds = np.zeros(sub_binary.shape, dtype=np.uint64)
seeds[seed1] = 1
seeds[seed2] = 2
for i in range(10):
seeds = mh.dilate(seeds)
ws = mh.cwatershed(grad, seeds)
ws[sub_binary == 0] = 0
# ws_relabeled = skimage.measure.label(ws.astype(np.uint8))
# ws_relabeled[sub_binary==0] = 0
# max_label = ws_relabeled.max()
# plt.figure()
# imshow(ws)
binary_mask = Util.threshold(ws, ws.max())
border = mh.labeled.border(ws, ws.max(), ws.max() - 1, Bc=mh.disk(2))
border[sub_binary_border == 1] = 0 # remove any "real" border pixels
# plt.figure()
# imshow(binary_mask)
# plt.figure()
# imshow(border)
# at this point, there can be multiple borders and labels
labeled_border = skimage.measure.label(border)
labeled_binary_mask = skimage.measure.label(binary_mask)
# .. and we are going to select only the largest
largest_border_label = Util.get_largest_label(
labeled_border.astype(np.uint16), True)
largest_binary_mask_label = Util.get_largest_label(
labeled_binary_mask.astype(np.uint16), True)
# .. filter out everything else
border[labeled_border != largest_border_label] = 0
binary_mask[labeled_binary_mask != largest_binary_mask_label] = 0
large_label = np.zeros(binary.shape, dtype=np.bool)
large_border = np.zeros(binary.shape, dtype=np.bool)
large_label[bbox[0]:bbox[1], bbox[2]:bbox[3]] = binary_mask
large_border[bbox[0]:bbox[1], bbox[2]:bbox[3]] = border
return large_label, large_border
def main(image, mask, threshold=150, bead_size=2, superpixel_size=4,
close_surface=False, close_disc_size=8, plot=False):
'''Converts an image stack with labelled cell surface to a cell
`volume` image
Parameters
----------
image: numpy.ndarray[Union[numpy.uint8, numpy.uint16]]
grayscale image in which beads should be detected (3D)
mask: numpy.ndarray[Union[numpy.int32, numpy.bool]]
binary or labeled image of cell segmentation (2D)
threshold: int, optional
intensity of bead (default: ``150``)
bead_size: int, optional
minimal size of bead (default: ``2``)
superpixel_size: int, optional
size of superpixels for searching the 3D position of a bead
close_surface: bool, optional
whether the interpolated surface should be morphologically closed
close_disc_size: int, optional
size in pixels of the disc used to morphologically close the
interpolated surface
plot: bool, optional
whether a plot should be generated (default: ``False``)
Returns
-------
jtmodules.generate_volume_image.Output
'''
n_slices = image.shape[-1]
logger.debug('input image has size %d in last dimension', n_slices)
logger.debug('mask beads inside cell')
beads_outside_cell = np.copy(image)
for iz in range(n_slices):
beads_outside_cell[mask > 0, iz] = 0
logger.debug('search for 3D position of beads outside cell')
slide = np.argmax(beads_outside_cell, axis=2)
slide[slide > np.percentile(slide[mask == 0], 20)] = 0
logger.debug('determine surface of slide')
slide_coordinates = array_to_coordinate_list(slide)
bottom_surface = fit_plane(subsample_coordinate_list(
slide_coordinates, 2000)
)
logger.debug('detect_beads in 2D')
mip = np.max(image, axis=-1)
try:
# TODO: use LOG filter???
beads, beads_centroids = detect_blobs(
image=mip, mask=np.invert(mask > 0), threshold=threshold,
min_area=bead_size
)
except:
logger.warn('detect_blobs failed, returning empty volume image')
volume_image = np.zeros(shape=mask.shape, dtype=image.dtype)
figure = str()
return Output(volume_image, figure)
n_beads = np.count_nonzero(beads_centroids)
logger.info('found %d beads on cells', n_beads)
if n_beads == 0:
logger.warn('empty volume image')
volume_image = np.zeros(shape=mask.shape, dtype=image.dtype)
else:
logger.debug('locate beads in 3D')
beads_coords_3D = locate_in_3D(
image=image, mask=beads_centroids,
bin_size=superpixel_size
)
logger.info('interpolate cell surface')
volume_image = interpolate_surface(
coords=beads_coords_3D,
output_shape=np.shape(image[:, :, 1]),
method='linear'
)
volume_image = volume_image.astype(image.dtype)
if (close_surface is True):
import mahotas as mh
logger.info('morphological closing of cell surface')
volume_image = mh.close(volume_image,
Bc=mh.disk(close_disc_size))
volume_image[mask == 0] = 0
if plot:
logger.debug('convert bottom surface plane to image for plotting')
bottom_surface_image = np.zeros(slide.shape, dtype=np.uint8)
for ix in range(slide.shape[0]):
for iy in range(slide.shape[1]):
bottom_surface_image[ix, iy] = plane(
ix, iy, bottom_surface.x)
logger.info('create plot')
from jtlib import plotting
plots = [
plotting.create_intensity_image_plot(
mip, 'ul', clip=True
),
plotting.create_intensity_image_plot(
bottom_surface_image, 'll', clip=True
),
plotting.create_intensity_image_plot(
volume_image, 'ur', clip=True
)
]
figure = plotting.create_figure(
plots, title='Convert stack to volume image'
)
else:
figure = str()
return Output(volume_image, figure)
