def NumpyDetectEdge(fname, color_filter=(1, 1, 1),
radius=510, center=(582, 791), show=False):
plate_circle = GetCircle(center, radius)
im = misc.imread(fname)
im_gray = np.dot(im, color_filter) / sum(color_filter)
imf = ndimage.gaussian_filter(im_gray, sigma=2)
imf = 256 - imf # invert color
imc = np.zeros(imf.shape, dtype=np.int32)
imc[plate_circle] = imf[plate_circle]
imc = imc[center[0]-radius:center[0]+radius, center[1]-radius:center[1]+radius]
T = 100
imc[np.where(imc < T)] = 0
rmax = pymorph.regmax(imc)
seeds, n_colonies = ndimage.label(rmax) # gives a unique integer number to each region and fills it with that number
count = 0
for i in xrange(n_colonies):
seed = np.where(seeds==i)
l = len(seed[0])
# filter seeds which are touching the perimeter (or 5 pixel away from it)
perimeter = False
for j in xrange(l):
if np.sqrt((radius-seed[0][j])**2 + (radius-seed[1][j])**2) > radius-5:
perimeter = True
if perimeter:
continue
seedx = [(x + center[0] - radius) for x in seed[0]]
seedy = [(y + center[1] - radius) for y in seed[1]]
meanx = np.mean(seedx)
meany = np.mean(seedy)
colony_circle = GetCircle((meanx, meany), 3)
colony_color = np.min(imf[colony_circle])
if colony_color >= 110:
count += 1
if colony_color < 100:
im[meanx, meany, :] = (255, 0, 0) # red
elif 100 < colony_color < 110:
im[meanx, meany, :] = (255, 255, 0) # yellow
elif 110 < colony_color < 120:
im[meanx, meany, :] = (0, 255, 0) # green
elif 120 < colony_color < 130:
im[meanx, meany, :] = (0, 255, 255) # cyan
elif 130 < colony_color < 140:
im[meanx, meany, :] = (0, 0, 255) # blue
elif 140 < colony_color < 150:
im[meanx, meany, :] = (255, 0, 255) # magenta
elif 150 < colony_color:
im[meanx, meany, :] = (255, 255, 255) # white
if show:
plt.imshow(im)
plt.show()
return count
def segment(img, T):
binimg = (img > T)
binimg = ndimage.median_filter(binimg, size=5)
dist = mahotas.distance(binimg)
dist = dist.astype(np.int32)
maxima = pymorph.regmax(dist)
maxima,_ = ndimage.label(maxima)
return mahotas.cwatershed(dist.max() - dist, maxima)
def labelimage(self, threshold=None):
"""Label regions in image corresponding to regional maxima"""
if threshold is None:
threshold = self.threshold
self.seeds = pymorph.regmax(self.filtered) * threshold
self.labels, self.nlabels = ndimage.label(self.seeds, structure=numpy.ones((3,3)))
def pymorph_objects(flnm, sigma):
raw_img = misc.imread('data/circles.png')
img = ndimage.gaussian_filter(raw_img, sigma)
thres = img > img.mean()
#find objects
peaks = pymorph.regmax(img)
found, n_found = ndimage.label(peaks)
com = ndimage.measurements.center_of_mass(peaks, found, xrange(1, n_found+1))
coords = com_text(com, n_found)
return '%d objects were found in %s.\nTheir centers of mass are located at \n %s' %(n_found, basename(flnm), coords)
def sobel_segment(img):
vsobel = np.array([
[1,2,1],
[0,0,0],
[-1,-2,-1]])
sobel = ndimage.convolve(img.astype(float),vsobel)**2
hsobel = vsobel.T
sobel += ndimage.convolve(img.astype(float), hsobel)**2
imgf = ndimage.gaussian_filter(img,2)
maxima,nmaxima = ndimage.label(pymorph.regmax(imgf))
overseg = mahotas.cwatershed(sobel.astype(np.uint32), maxima)
return overseg
def count_nuclei(filename): full = filename #nome immagine da analizzare empty = '/home/luca/isac/black-cut.bmp' #immagine vuota di riferimento img = '/home/luca/isac/tmp/'+filename+'.jpg' #nome del file da creare con imagemagick imagemagick= "compare -metric AE -fuzz 8% " +empty+" "+full+" "+ " -highlight-color White -lowlight-color Black " + img #good values 5-6-7-8% os.system(imagemagick) #esegue il comando di imagemagick creando il file ccnx.jpg per l'immagine corrente ccn = mahotas.imread(img) #carica l'immagine creata con imagemagick ccnf = ndimage.gaussian_filter(ccn, 10) #applica un filtro gaussiano con SD=10 all'immagine rmax = pymorph.regmax(ccnf) #trova i massimi locali nell'immagine seeds,nr_nuclei=ndimage.label(rmax) #conta i massimi locali dell'immagine filtrata return nr_nuclei
def GradBasedSegmentation(im):
blur = nd.gaussian_filter(im, 16)
rmax = pymorph.regmax(blur)
T = mahotas.thresholding.otsu(blur)
bImg0 = im > T
# bImg01=nd.binary_closing(bImg0,iterations=2)
bImg01 = pymorph.close(bImg0, pymorph.sedisk(3))
bImg = pymorph.open(bImg01, pymorph.sedisk(4))
# bImg=nd.binary_opening(bImg01,iterations=3)
b = pymorph.edgeoff(bImg)
d = distanceTranform(b)
seeds, nr_nuclei = nd.label(rmax)
lab = mahotas.cwatershed(d, seeds)
return lab
def GradBasedSegmentation(im):
blur=nd.gaussian_filter(im, 16)
rmax = pymorph.regmax(blur)
T = mahotas.thresholding.otsu(blur)
bImg0=im>T
#bImg01=nd.binary_closing(bImg0,iterations=2)
bImg01=pymorph.close(bImg0, pymorph.sedisk(3))
bImg=pymorph.open(bImg01, pymorph.sedisk(4))
#bImg=nd.binary_opening(bImg01,iterations=3)
b=pymorph.edgeoff(bImg)
d=distanceTranform(b)
seeds,nr_nuclei = nd.label(rmax)
lab=mahotas.cwatershed(d,seeds)
return lab
def colorscheme(url):
im=image_read(url)
pylab.imshow(im)
dnaf = ndimage.gaussian_filter(im, 1)
rmax = pymorph.regmax(dnaf)
seeds,nr_nuclei = ndimage.label(rmax)
T = mahotas.thresholding.otsu(dnaf)
dist = ndimage.distance_transform_edt(dnaf > T)
dist = dist.max() - dist
dist -= dist.min()
dist = dist/float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
nuclei = pymorph.cwatershed(dist, seeds)
whole = mahotas.segmentation.gvoronoi(nuclei)
im0=pylab.imshow(whole)
pylab.show()
def maxima(img):
"""
Return image local maxima
"""
import pymorph
from image import Transf
# Image has to be 'int' [0:255] to use in 'pymorph'
img = Transf.float2uint(img)
# Search for image maxima
maxs = pymorph.regmax(img)
# A closing step is used "clue" near maxima
elem_strct = ndi.generate_binary_structure(2,2)
maxs = ndi.binary_closing(maxs,elem_strct)
return maxs
def watershed(img):
# Diminui ruidos
imgf = ndimage.gaussian_filter(img, 16)
mahotas.imsave("dnaf.jpeg", imgf)
rmax = pymorph.regmax(imgf)
T = mahotas.thresholding.otsu(imgf)
dist = ndimage.distance_transform_edt(imgf > T)
dist = dist.max() - dist
dist -= dist.min()
dist = dist / float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
mahotas.imsave("dist.jpeg", dist)
seeds, nr_nuclei = ndimage.label(rmax)
nuclei = pymorph.cwatershed(dist, seeds)
mahotas.imsave("watershed.jpeg", nuclei)
def start(self):
"""Segment frame.
The returned value is a labeled uint16 image.
"""
# Preprocessing: subtract minimum pixel value.
I = self._image - self._image.min()
# 'Bandpass' filtering.
I_s = ndimage.filters.gaussian_filter(I, self._sigma_s) # Foreground.
I_b = ndimage.filters.gaussian_filter(I, self._sigma_b) # Background.
I_bp = I_s - self._alpha * I_b
# Thresholding: create binary image.
I_bin = (I_bp > self._tau)
# Hole filling.
I_bin = ndimage.binary_fill_holes(I_bin > 0)
I_cells = ndimage.label(I_bin)[0]
# Avoid merging nearby cells using watershed.
if self._watershed:
# Distance transfrom on which to apply the watershed algorithm.
I_dist = ndimage.distance_transform_edt(I_bin)
I_dist = I_dist/float(I_dist.max()) * 255
I_dist = I_dist.astype(np.uint8)
# Find markers for the watershed algorithm.
# Reduce false positive using Gaussian smoothing.
I_mask = ndimage.filters.gaussian_filter(I_dist, 8)*I_bin
rmax = pymorph.regmax(I_mask)
I_markers, num_markers = ndimage.label(rmax)
I_dist = I_dist.max() - I_dist # Cells are now the basins.
I_cells = pymorph.cwatershed(I_dist, I_markers)
# Remove cells with area less than threshold.
if self._a_min:
for label in np.unique(I_cells)[1:]:
if (I_cells == label).sum() < self._a_min:
I_cells[I_cells == label] = 0
# Remove cells with summed intensity less than threshold.
if self._s_min:
for label in np.nditer(np.unique(I_cells)[1:]):
if I_bp[I_cells == label].sum() < self._s_min:
I_cells[I_cells == label] = 0
return I_cells.astype('uint16') # This data type is used by ISBI.
def watershed_segment(img, mode='direct', thresholding=None, min_obj_size=None, **kwargs):
'''
segment_watershed(img, mode='direct', thresholding=None, min_obj_size=None, **kwargs)
Segment using traditional watershed
Parameters
----------
* img: a pyslic.Image. The algorithm operates on the dna channel.
* mode: 'direct' or 'gradient': whether to use the image or the gradient of the image
* thresholding: how to threshold the smoothed image (default: None, no thresholding)
* min_obj_size: minimum object size. This is slightly different than post-filtering for minimum
object size as it fill those holes with a second watershed pass as opposed to having
an image with holes
* smoothing: whether to smooth (default: True)
* smooth_gamma: Size of Gaussian for blurring, in pixels (default: 12)
'''
assert mode in ('direct','gradient'), "segment_watershed: mode '%s' not understood" % mode
with loadedimage(img):
dna = img.get('dna')
if kwargs.get('smoothing',True):
dnaf = ndimage.gaussian_filter(dna, kwargs.get('smooth_gamma',12))
else:
dnaf = dna
rmax = pymorph.regmax(dnaf)
rmax_L,_ = ndimage.label(rmax)
if mode == 'direct':
watershed_img = dna.max()-dna
elif mode == 'gradient':
dnag = pymorph.gradm(dna)
watershed_img = dnag.max()-dnag
water = mahotas.cwatershed(watershed_img,rmax_L)
if thresholding is not None:
T = threshold(dnaf,thresholding)
water *= (dnaf >= T)
if min_obj_size is not None:
oid = 1
while oid <= water.max():
if (water == oid).sum() < min_obj_size:
water[water == oid] =0
water[water > oid] -= 1
else:
oid += 1
return water
def effect(url):
im=image_read(url)
pylab.imshow(im)
dnaf = ndimage.gaussian_filter(im, 1)
rmax = pymorph.regmax(dnaf)
T = mahotas.thresholding.otsu(dnaf)
seeds,nr_nuclei = ndimage.label(rmax)
T = mahotas.thresholding.otsu(dnaf)
T = mahotas.thresholding.otsu(dnaf)
dist = ndimage.distance_transform_edt(dnaf > T)
dist = dist.max() - dist
dist -= dist.min()
dist = dist/float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
nuclei = pymorph.cwatershed(dist, seeds)
pylab.imshow(nuclei)
# pylab.save("out.png")
pylab.show()
def MNUC(datatype, maxrange, outputfile, outputfiletype):
h = open(outputfile, outputfiletype)
TC = 0
for i in range(0, maxrange + 1):
A = datatype[i][0]
rmax = pymorph.regmax(A)
seeds, nr_nuclei = ndimage.label(rmax)
T = mahotas.thresholding.otsu(A)
C = A.copy()
if T < 1:
C[ C <= T ] = 0
C[ C > T ] = 1
else:
C[ C < T ] = 0
C[ C >= T ] = 1
filled = scipy.ndimage.morphology.binary_fill_holes(C)
filled = filled.astype(np.uint8)
dist = ndimage.distance_transform_edt(filled > T)
dist = dist.max() - dist
dist -= dist.min()
dist = dist/float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
nuclei = pymorph.cwatershed(dist, seeds)
find = mahotas.label(nuclei)
for n in range(0, find[1] + 1):
CD = np.where(find[0] == n)
XY1 = np.vstack((CD[0], CD[1]))
edges = filter.canny(XY1, sigma=1)
edges = edges.astype(np.uint8)
edges = np.where(edges == 1)
TC += len(CD[0])
XY1 = np.vstack((CD[0], CD[1], [i*5]*len(CD[0]), [n]*len(CD[0])))
for p in range(0, len(XY1[0])):
for yel in range(0, len(XY1)):
h.write(str(XY1[yel][p]) + '\t')
h.write('\n')
h.write(str(TC) + '\n')
h.write('.' + '\n')
h.close()
#
#
# Predictions with regmax method
blur_imgH = scipy.ndimage.gaussian_filter(mito_prob, 16)
mito_pred3 = blur_imgH<90
blur_imgH = blur_imgH.astype(np.uint8)
mito_pred3 = mahotas.erode(mito_pred3, disc)
mito_pred3 = mito_pred3.astype(np.uint8)
# labeled, nr_objects = scipy.ndimage.label(mito_pred3)
# print nr_objects
# labeled = labeled.astype(np.uint8)
rmax = pymorph.regmax(blur_imgH)
pylab.imshow(pymorph.overlay(mito_prob, rmax))
pylab.gray()
pylab.show()
seeds,nr_nuclei = scipy.ndimage.label(rmax)
print nr_nuclei
pylab.imshow(mito_pred3)
pylab.show()
dist = scipy.ndimage.distance_transform_edt(mito_pred3)
dist = dist.max() - dist
dist-=dist.min()
dist = dist/float(dist.ptp())*255
dist = dist.astype(np.uint8)
pylab.imshow(dist)
pylab.gray()
pylab.show()
def SearchMarker(In, image_filtree, marker_param, mask):
"""Algorithme principale de recherche de marqueur et segmentation
:In: image d'entree
:image_filtree: image filtree par MeanShift
:marker_param: choix de la methode de recherche (1,2 ou 3)
:mask: masque de non interet
:returns: image label, image pour affichee
"""
if image_filtree.nChannels == 1:
tmp = cv.CreateImage(cv.GetSize(image_filtree), cv.IPL_DEPTH_8U, 3)
cv.CvtColor(image_filtree, tmp, cv.CV_GRAY2BGR)
image_filtree = cv.CloneImage(tmp)
del tmp
fg = cv.CloneImage(In)
objets = None
# Image distance entre de watershed
markers = cv.CreateImage(cv.GetSize(In), cv.IPL_DEPTH_32S, 1)
img = cv.CloneImage(image_filtree)
cv.Zero(img)
edge = Detection_contour(In)
cont = cv2array(edge)[:, :, 0]
cv.Not(edge, edge)
dist_map = cv.CreateImage(cv.GetSize(In), cv.IPL_DEPTH_32F, 1)
cv.DistTransform(edge, dist_map, cv.CV_DIST_L2, cv.CV_DIST_MASK_5)
dist_map_8bit = cv.CloneImage(edge)
cv.Zero(dist_map_8bit)
cv.ConvertScale(dist_map, dist_map, 3000.0, 0.0)
cv.Pow(dist_map, dist_map, 0.3)
cv.ConvertScale(dist_map, dist_map_8bit)
cv.CvtColor(dist_map_8bit, img, cv.CV_GRAY2BGR) #
cv.AddWeighted(image_filtree, 0.3, img, 0.7, 1, img) #
cv.CvtColor(img, dist_map_8bit, cv.CV_BGR2GRAY) #
# Foreground by regional maxima: detection marqueurs
if marker_param == "1" or marker_param == "3":
print("Recherche max. regionaux...")
I = cv2array(dist_map_8bit)[:, :, 0]
If = ndimage.gaussian_filter(I, 5)
rmax = pymorph.regmax(If)
rmax = pymorph.close_holes(rmax) * 255
#import ipdb;ipdb.set_trace()
bool_fg = array2cv(rmax.astype(np.uint8))
cv.ConvertScale(bool_fg, fg)
if marker_param == "1":
print("Recherche squelette...")
from mamba import *
from mambaComposed import *
from MambaTools import *
percent_edge = np.sum(cont) / (edge.width * edge.height)
initial_shape = (In.height, In.width)
im1 = fillImageWithIplimage(In)
imWrk1 = imageMb(im1)
blobsMarkers = imageMb(im1, 1)
imWrk3 = imageMb(im1, 32)
#backgroundMarker = imageMb(im1, 1)
#finalMarkers = imageMb(im1, 1)
print("taille se %s" % int(15.0 * 6 / percent_edge + 1))
if In.height < 700:
alternateFilter(im1, imWrk1,
int(15.0 * 6 / percent_edge) + 1, True)
elif In.height < 1400:
alternateFilter(im1, imWrk1,
int(30.0 * 6 / percent_edge) + 1, True)
else:
alternateFilter(im1, imWrk1,
int(60.0 * 6 / percent_edge) + 1, True)
minima(imWrk1, blobsMarkers)
thinD(blobsMarkers, blobsMarkers)
nbStones = label(blobsMarkers, imWrk3)
bg_array = getArrayFromImage(imWrk3)[
0: initial_shape[0],
0: initial_shape[1]]
tmp_array = (bg_array > 0) * 255
bg_ = array2cv(tmp_array.astype(np.uint8))
bg = cv.CloneImage(dist_map_8bit)
cv.ConvertScale(bg_, bg)
cv.Or(fg, bg, fg)
cv.ConvertScale(fg, markers)
# Watershed
print("Watershed...")
storage = cv.CreateMemStorage(0)
contours = cv.FindContours(fg, storage,
cv.CV_RETR_CCOMP,
cv.CV_CHAIN_APPROX_SIMPLE)
def contour_iterator(contour):
while contour:
yield contour
contour = contour.h_next()
cv.Zero(markers)
comp_count = 0
for c in contour_iterator(contours):
cv.DrawContours(markers,
c,
cv.ScalarAll(comp_count + 1),
cv.ScalarAll(comp_count + 1),
-1,
-1,
8)
comp_count += 1
cv.Watershed(img, markers)
if img.nChannels == 3:
cv.CvtColor(In, img, cv.CV_GRAY2BGR)
elif img.nChannels == 1:
img = cv.CloneImage(image_filtree)
cv.CvtColor(In, img, cv.CV_GRAY2BGR)
else:
print("nChannels >< 3 or 1")
cv.CvtColor(fg, img, cv.CV_GRAY2BGR)
cv.CvtColor(In, img, cv.CV_GRAY2BGR)
#bug
wshed = Affichage_watershed(markers, img, fg)
wshed_lbl = cv2array(markers)[:, :, 0]
if marker_param == "1" or marker_param == "2":
objets = cv2array(bg)[:, :, 0]
objets = objets > 1
print("Keep objects not in background ...")
cmpt = 0
label_map = wshed_lbl * 0
for label in np.unique(wshed_lbl):
if np.sum(objets * (wshed_lbl == label)) > 0:
label_map[wshed_lbl == label] = 0
else:
cmpt += 1
label_map[wshed_lbl == label] = cmpt
else:
label_map = wshed_lbl
label_map = Enleve_bord(label_map, mask)
return label_map, wshed
import numpy
import copy
import skimage.filter as skif
import skimage.morphology as morph
import matplotlib.pyplot as mpl
import skimage.feature as feature
import pylab
import mahotas
import pymorph
from skimage.filter import canny
### Circles ###
raw_circle = misc.imread('circles.png')
img_circle = ndimage.gaussian_filter(raw_circle, 15)
rmax_circle = pymorph.regmax(img_circle)
pylab.imshow(pymorph.overlay(img_circle, rmax_circle))
pylab.show()
seeds_circle, nr_circles = ndimage.label(rmax_circle)
print "Number of objects: " + str(nr_circles)
centers_circles = ndimage.center_of_mass(rmax_circle, seeds_circle,
range(1, nr_circles + 1, 1))
print "Centers of gravity: " + str(centers_circles)
### Objects ###
raw_objects = misc.imread('objects.png')
img_objects = ndimage.gaussian_filter(raw_objects, 3)
thres_objects = img_objects > img_objects.mean()
pylab.imshow(thres_objects)
pylab.show()
pass
def writeimg(img, name):
pass
else:
def savejet(img, name):
img = (img - img.min()) / float(img.ptp()) * 255
img = img.astype(np.uint8)
readmagick.writeimg((cm.jet(img)[:, :, :3] * 255).astype(np.uint8),
name)
writeimg = readmagick.writeimg
dnaf = ndimage.gaussian_filter(dna, 8)
rmax = pymorph.regmax(dnaf)
pylab.imshow(pymorph.overlay(dna, rmax))
pylab.show()
writeimg(pymorph.overlay(dna, rmax), 'dnaf-rmax-overlay.jpeg')
savejet(dnaf, 'dnaf-8.jpeg')
dnaf = ndimage.gaussian_filter(dna, 16)
rmax = pymorph.regmax(dnaf)
pylab.imshow(pymorph.overlay(dna, rmax))
pylab.show()
writeimg(pymorph.overlay(dna, rmax), 'dnaf-16-rmax-overlay.jpeg')
seeds, nr_nuclei = ndimage.label(rmax)
print nr_nuclei
T = pit.thresholding.otsu(dnaf)
def _segment(self, I, first):
"""Return the segmented frame 'I'.
If 'first is True, then this is the first segmentation iteration,
otherwise the second.
The returned value is a labeled image of type uint16, in order to be
compatible with ISBI's tool.
"""
# Compute global threshold.
otsu_thresh = mh.thresholding.otsu(I.astype('uint16'))
# Threshold using global and local thresholds.
fnc = fnc_class(I.shape)
I_bin = ndimage.filters.generic_filter(I, fnc.filter, size=self._r,
extra_arguments=(I, self._min_var,
otsu_thresh))
I_med = ndimage.filters.median_filter(I_bin, size=self._r_med)
# Remove cells which are too small (leftovers).
labeled = mh.label(I_med)[0]
sizes = mh.labeled.labeled_size(labeled)
too_small = np.where(sizes < self._a_min)
I_cleanup = mh.labeled.remove_regions(labeled, too_small)
I_cleanup = mh.labeled.relabel(I_cleanup)[0]
# Fill holes.
I_holes = ndimage.morphology.binary_fill_holes(I_cleanup > 0)
# Binary closing.
if first and self._r1:
# First iteration.
I_morph = morph.binary_closing(I_holes, morph.disk(self._r1))
elif not first and self._r2:
# Second iteration.
I_morph = morph.binary_closing(I_holes, morph.disk(self._r2))
else:
# No binary closing.
I_morph = I_holes
# Fill yet to be filled holes.
labels = measure.label(I_morph)
labelCount = np.bincount(labels.ravel())
background = np.argmax(labelCount)
I_morph[labels != background] = True
# Separate touching cells using watershed.
# Distance transfrom on which to apply the watershed algorithm.
I_dist = ndimage.distance_transform_edt(I_morph)
I_dist = I_dist/float(I_dist.max()) * 255
I_dist = I_dist.astype(np.uint8)
# Find markers for the watershed algorithm.
# Reduce false positive using Gaussian smoothing.
I_mask = ndimage.filters.gaussian_filter(I_dist, 8)*I_morph
rmax = pymorph.regmax(I_mask)
I_markers, _ = ndimage.label(rmax)
I_dist = I_dist.max() - I_dist # Cells are now the basins.
I_label = pymorph.cwatershed(I_dist, I_markers)
if self._debug:
plt.subplot(2, 4, 1)
plt.imshow(I)
plt.title('Original Image')
plt.subplot(2, 4, 2)
plt.imshow(I_bin)
plt.title('After Thresholding')
plt.subplot(2, 4, 3)
plt.imshow(I_med)
plt.title('After Median Filter')
plt.subplot(2, 4, 4)
plt.imshow(I_cleanup)
plt.title('After Cleanup')
plt.subplot(2, 4, 5)
plt.imshow(I_holes)
plt.title('After Hole Filling')
plt.subplot(2, 4, 6)
plt.imshow(I_morph)
plt.title('After Closing')
plt.subplot(2, 4, 7)
plt.imshow(I_label)
plt.title('Labeled Image')
plt.show()
return I_label.astype('uint16')
def main():
# Load and show original images
pylab.gray() # set gray scale mode
print
print "0. Reading and formatting images..."
images = {f: loadAndFormat(f) for f in IMAGE_FILES}
for f in IMAGE_FILES:
mShow(images[f])
###########################
# -----> Thresholding
print
print "1. Thresholding images..."
thresholdedImages = {f: getThresholdedImage(images[f]) for f in IMAGE_FILES}
for name in IMAGE_FILES:
mShow(thresholdedImages[name])
###########################
# -----> Count objects
# 1st attempt: label the thresholded image from task 1
print
print "2. Object counting"
pylab.jet() # back to color mode
print "\t1st approach: Label thresholded images"
for name in IMAGE_FILES:
labeled, nrRegions = ndimage.label(thresholdedImages[name])
print "\t" + name + ": " + str(nrRegions)
mShow(labeled)
# 2nd attempt: Changing threshold level
print
print "\t2nd approach: Tuned thresholds"
# For 'objects.png' some objects are very small (e.g.: screw) or
# have many shades (e.g.: spoon) which makes them disappear or appear
# fragmented after thresholding.
# The advantage of this image is that the background is very dark,
# so we can try using a lower threshold to make all shapes more definite
objImage = images['objects.png']
T = mahotas.thresholding.otsu(objImage)
thresholdedImage = objImage > T * 0.7
# Looks better, but...
labeled, nrRegions = ndimage.label(thresholdedImage)
print '\tobjects.png' + ": " + str(nrRegions)
# it returns 18!
# 3rd attempt: Smoothing before thresholding
print
print "\t3rd approach: Smoothing + Tuned threshold"
# Let's apply some Gaussian smoothing AND a lower threshold
smoothImage = ndimage.gaussian_filter(objImage, 3)
T = mahotas.thresholding.otsu(smoothImage)
thresholdedImage = smoothImage > T * 0.7
labeled, nrRegions = ndimage.label(thresholdedImage)
print '\tobjects.png' + ": " + str(nrRegions)
# it worked! Let's save the labeled images for later
# (we will use them for center calculation)
labeledImages = {}
labeledImages['objects.png'] = (labeled, nrRegions)
mShow(labeled)
# Let's see if this approach works on the other images
for name in ['circles.png', 'peppers.png']:
img = images[name]
smoothImage = ndimage.gaussian_filter(img, 3)
T = mahotas.thresholding.otsu(smoothImage)
thresholdedImage = smoothImage > T * 0.7
labeled, nrRegions = ndimage.label(thresholdedImage)
print '\t' + name + ": " + str(nrRegions)
# Again no luck with the circles!
# (We will take a closer look at the peppers later)
# 4th attempt:
# 'circles.png': The problem is that some circles appear "glued together".
# Let's try another technique:
# - smoothing the picture with a Gaussian filter
# - then searching for local maxima and counting regions
# (smoothing avoids having many scatter maxima and a higher level
# must be used than in the previous attempt)
# - use watershed with the maxima as seeds over the thresholded image
# to complete the labelling of circles
print
print "\t4th approach: Smoothing + Local maxima + Watershed"
smoothImage = ndimage.gaussian_filter(images['circles.png'], 10)
localmaxImage = pymorph.regmax(smoothImage)
# A distance transform must be applied before doing the watershed
dist = ndimage.distance_transform_edt(thresholdedImages['circles.png'])
dist = dist.max() - dist
dist -= dist.min()
dist = dist / float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
seeds, nrRegions = ndimage.label(localmaxImage)
labeled = pymorph.cwatershed(dist, seeds)
print "\t" + 'circles.png' + ": " + str(nrRegions)
# worked right only for 'circles.png' !
labeledImages['circles.png'] = (labeled, nrRegions)
mShow(labeled)
print
print "\t5th approach: Smoothing + Multi-threshold +" +\
" Morphology labeling + Size filtering"
# 5th approach (only peppers left!)
imagePeppers = images['peppers.png']
# Problems with peppers are:
# - very different colours, they cause thresholding to work poorly
# - each pepper has some brighter parts which are detected as local maxima
# We propose to address those issues as follows:
# - gaussian filter to smooth regions of light or shadow within each pepper
smoothImage = ndimage.gaussian_filter(imagePeppers, 2)
# - instead of thresholding to create a binary image,
# create multiple thresholds to separate the most frequent colors.
# In this case, 3 thresholds will be enough
mthrImagePeppers = multiThreshold(smoothImage, 3)
# - ndimage.label didn't give good results, we try another
# labelling algorithm
from skimage import morphology
labeled = morphology.label(mthrImagePeppers)
nrRegions = np.max(labeled) + 1
print "\t\tTotal number of regions"
print "\t\t\t" + 'peppers.png' + ": " + str(nrRegions)
# - after counting regions, filter to keep only the sufficiently big ones
filtered, nrRegions = filterRegions(labeled, 0.05)
print "\t\tBig enough regions"
print "\t\t\t" + 'peppers.png' + ": " + str(nrRegions)
labeledImages['peppers.png'] = (filtered, nrRegions)
mShow(filtered)
###########################
# -----> Find center points
print
print "3. Centers for objects"
for img in IMAGE_FILES:
labeledImage, nr_objects = labeledImages[img]
CenterOfMass = ndimage.measurements.center_of_mass
labels = range(1, nr_objects + 1)
centers = CenterOfMass(labeledImage, labeledImage, labels)
centers = [(int(round(x)), int(round(y))) for (x, y) in centers]
print '\t' + img + ": " + str(centers)
# Isa = np.rollaxis(np.dstack(tiffile.imread(i)), 2, 0)
Is = np.rollaxis(np.dstack([tiffile.imread(i) for i in img_files]), 2, 0)
movie = Is
Is = Is[:,::2,::2].astype('float64')
# Calculate the maximum across all frames
accum = np.log(Is.max(axis=0))
import skimage.segmentation, skimage.feature, skimage.morphology.watershed
import mahotas, pymorph
I_show = accum.copy()
#increase contrast
import ImageEnhance
I_contrast = pow(I_show.astype("uint16"), 3)
#find regional maxima
regionalMax = pymorph.regmax((I_contrast).astype("uint16"))
#find seeds and cell number
seeds,numCells = ndimage.label(regionalMax)
print numCells
#edge detection
T = mahotas.thresholding.otsu(I_contrast.astype("uint16"))
dist = ndimage.distance_transform_edt(I_contrast.astype("uint16") > T)
dist = dist.max() - dist
dist -= dist.min()
dist = dist/float(dist.ptp()) * 255
dist = dist.astype(np.uint8)
#watershed
I_mask = pymorph.cwatershed(dist, seeds)
frames, x, y = Is.shape
import numpy
import copy
import skimage.filter as skif
import skimage.morphology as morph
import matplotlib.pyplot as mpl
import skimage.feature as feature
import pylab
import mahotas
import pymorph
from skimage.filter import canny
### Circles ###
raw_circle = misc.imread('circles.png')
img_circle = ndimage.gaussian_filter(raw_circle, 15)
rmax_circle = pymorph.regmax(img_circle)
pylab.imshow(pymorph.overlay(img_circle, rmax_circle))
pylab.show()
seeds_circle, nr_circles = ndimage.label(rmax_circle)
print "Number of objects: " + str(nr_circles)
centers_circles = ndimage.center_of_mass(rmax_circle, seeds_circle, range(1, nr_circles + 1, 1))
print "Centers of gravity: " + str(centers_circles)
### Objects ###
raw_objects = misc.imread('objects.png')
img_objects = ndimage.gaussian_filter(raw_objects, 3)
thres_objects = img_objects > img_objects.mean()
pylab.imshow(thres_objects)
pylab.show()
# regmax is resulting in an error. Looks like this is working
pylab.imshow(dnaf > T) pylab.show() # Deal with merged/touching nuclei labeled,nr_objects = ndimage.label(dnaf > T) print nr_objects # prints 18 pylab.imshow(labeled) pylab.jet() # resets to jet from gray-scale pylab.show() ############ STEP TWO -- Segmenting Image/Finding seeds # Smooth image->find regional maxima->use maxima as seeds for watershed # First try: dnaf = ndimage.gaussian_filter(dna, 8) rmax = pymorph.regmax(dnaf) pylab.imshow(pymorph.overlay(dna, rmax)) # Overlay returns a color image with gray level component in first argument, second arg is red pylab.show() # Second try - Increase sigma: dnaf = ndimage.gaussian_filter(dna, 16) rmax = pymorph.regmax(dnaf) pylab.imshow(pymorph.overlay(dna, rmax)) seeds,nr_nuclei = ndimage.label(rmax) print nr_nuclei # prints 22 # Watershed to distance transform of threshold T = mahotas.thresholding.otsu(dnaf) dist = ndimage.distance_transform_edt(dnaf > T) dist = dist.max() - dist
def NumpyDetectEdge(fname,
color_filter=(1, 1, 1),
radius=510,
center=(582, 791),
show=False):
plate_circle = GetCircle(center, radius)
im = misc.imread(fname)
im_gray = np.dot(im, color_filter) / sum(color_filter)
imf = ndimage.gaussian_filter(im_gray, sigma=2)
imf = 256 - imf # invert color
imc = np.zeros(imf.shape, dtype=np.int32)
imc[plate_circle] = imf[plate_circle]
imc = imc[center[0] - radius:center[0] + radius,
center[1] - radius:center[1] + radius]
T = 100
imc[np.where(imc < T)] = 0
rmax = pymorph.regmax(imc)
seeds, n_colonies = ndimage.label(
rmax
) # gives a unique integer number to each region and fills it with that number
count = 0
for i in xrange(n_colonies):
seed = np.where(seeds == i)
l = len(seed[0])
# filter seeds which are touching the perimeter (or 5 pixel away from it)
perimeter = False
for j in xrange(l):
if np.sqrt((radius - seed[0][j])**2 +
(radius - seed[1][j])**2) > radius - 5:
perimeter = True
if perimeter:
continue
seedx = [(x + center[0] - radius) for x in seed[0]]
seedy = [(y + center[1] - radius) for y in seed[1]]
meanx = np.mean(seedx)
meany = np.mean(seedy)
colony_circle = GetCircle((meanx, meany), 3)
colony_color = np.min(imf[colony_circle])
if colony_color >= 110:
count += 1
if colony_color < 100:
im[meanx, meany, :] = (255, 0, 0) # red
elif 100 < colony_color < 110:
im[meanx, meany, :] = (255, 255, 0) # yellow
elif 110 < colony_color < 120:
im[meanx, meany, :] = (0, 255, 0) # green
elif 120 < colony_color < 130:
im[meanx, meany, :] = (0, 255, 255) # cyan
elif 130 < colony_color < 140:
im[meanx, meany, :] = (0, 0, 255) # blue
elif 140 < colony_color < 150:
im[meanx, meany, :] = (255, 0, 255) # magenta
elif 150 < colony_color:
im[meanx, meany, :] = (255, 255, 255) # white
if show:
plt.imshow(im)
plt.show()
return count
