def ThresholdMaxEntropy(imp0):
"""Thresholds image and returns thresholded image, merge code still quite clumsy but functional"""
imp0 = IJ.getImage()
impthres = imp0.duplicate()
imp01 = ImagePlus("Channel1", ChannelSplitter.getChannel(imp0, 1))
imp02 = ImagePlus("Channel2", ChannelSplitter.getChannel(imp0, 2))
imp001 = imp01.duplicate()
imp002 = imp02.duplicate()
IJ.setAutoThreshold(imp001, "MaxEntropy dark")
IJ.run(imp001, "Convert to Mask", "")
IJ.run(imp001, "Divide...", "value=255")
IJ.setAutoThreshold(imp002, "MaxEntropy dark")
IJ.run(imp002, "Convert to Mask", "")
IJ.run(imp002, "Divide...", "value=255")
ic = ImageCalculator()
imp0001 = ic.run("Multiply create", imp01, imp001)
ic2 = ImageCalculator()
imp0002 = ic2.run("Multiply create", imp02, imp002)
imp0001.copy()
impthres.setC(1)
impthres.paste()
imp0002.copy()
impthres.setC(2)
impthres.paste()
imp01.close()
imp02.close()
imp001.close()
imp002.close()
imp0001.close()
imp0002.close()
return impthres
def getCroppedChannels(self, imp, cell):
splitter = ChannelSplitter()
imp.setRoi(None)
if cell.mode3D:
cropRoi = cell.getCropRoi()
else:
cropRoi = cell.roi
if cropRoi is None:
return None
crop = cropRoi.getBounds()
channels = []
for c in range(1, imp.getNChannels() + 1):
slices = ImageStack(crop.width, crop.height)
channel = splitter.getChannel(imp, c)
for z in range(1, channel.getSize() + 1):
zslice = channel.getProcessor(z)
zslice.setRoi(cropRoi)
nslice = zslice.crop()
if cell.mode3D:
oroi = cell.slices[z - 1].roi
else:
oroi = cell.roi
if oroi is not None:
roi = oroi.clone()
bounds = roi.getBounds()
roi.setLocation(bounds.x - crop.x, bounds.y - crop.y)
nslice.setColor(Color.black)
nslice.fillOutside(roi)
slices.addSlice(nslice)
channels.append(ImagePlus("Channel %i" % c, slices))
return channels
def split_channels_blue(imp):
#Create blue channel
original_blue = ChannelSplitter.getChannel(imp, 3)
original_blue_IP = ImagePlus(filename, original_blue)
fs = FileSaver(original_blue_IP)
folder = "/Users/gceleste/Desktop/test/channels"
filepath = folder + "/" + "{}_bluechannel.tif".format(filename)
fs.saveAsTiff(filepath)
#Open blue channel image.
blue = IJ.open(filepath)
#blue_IP = ImagePlus(filename, blue)
IJ.run(
"3D Objects Counter",
"threshold=100 slice=1 min.=50 max.=1447680 exclude_objects_on_edges objects summary"
)
#Save blue object map
blue_map = IJ.getImage()
fs = FileSaver(blue_map)
folder = "/Users/gceleste/Desktop/test/object maps"
filepath = folder + "/" + "{}_objectmap(blue).jpg".format(filename)
fs.saveAsJpeg(filepath)
#Close blue channel image.
blue_map.close()
blue = IJ.getImage()
blue.close()
def split_channels_red(imp):
#Create red channel
original_red = ChannelSplitter.getChannel(imp, 1)
original_red_IP = ImagePlus(filename, original_red)
fs = FileSaver(original_red_IP)
folder = "/Users/gceleste/Desktop/test/channels"
filepath = folder + "/" + "{}_redchannel.tif".format(filename)
fs.saveAsTiff(filepath)
#Open red channel image.
red = IJ.open(filepath)
#red_IP = ImagePlus(filename, red)
IJ.run(
"3D Objects Counter",
"threshold=130 slice=1 min.=50 max.=1447680 exclude_objects_on_edges objects summary"
)
#Save red object map.
red_map = IJ.getImage()
fs = FileSaver(red_map)
folder = "/Users/gceleste/Desktop/test/object maps"
filepath = folder + "/" + "{}_objectmap(red).jpg".format(filename)
fs.saveAsJpeg(filepath)
#Close red channel images.
red_map.close()
red = IJ.getImage()
red.close()
def previewImage(self, imp):
roi = imp.getRoi()
splitter = ChannelSplitter()
channels = []
for c in range(1, imp.getNChannels() + 1):
channel = ImagePlus("Channel %i" % c, splitter.getChannel(imp, c))
projector = ZProjector(channel)
projector.setMethod(ZProjector.MAX_METHOD)
projector.doProjection()
channels.append(projector.getProjection())
image = RGBStackMerge.mergeChannels(channels, False)
image.title = imp.title + " MAX Intensity"
image.luts = imp.luts
imp.setRoi(roi)
return image
def getPreview(image):
enhancer = ContrastEnhancer()
projector = ZProjector()
splitter = ChannelSplitter()
imp1 = ImagePlus("CH1", )
width, height, channels, slices, frames = image.getDimensions()
chimps = []
for ch in range(1, channels + 1):
projector = ZProjector(ImagePlus("C%i" % ch, splitter.getChannel(image, ch)))
projector.setMethod(ZProjector.MAX_METHOD)
projector.doProjection()
proj = projector.getProjection()
enhancer.equalize(proj)
chimps.append(proj)
return RGBStackMerge.mergeChannels(chimps, False)
f for f in os.listdir(os.path.join(inputPath)) if f.lower()[0].isalnum()
]
for sample in imagelist:
rmi.reset()
IJ.run('Close All')
sample_label = sample.split('.')[0]
outputFile = os.path.join(outputPath, sample_label + '.zip')
if not os.path.exists(outputFile): #Check that file does not yet exists
print 'Segmenting ' + sample_label
#Load images
imp = IJ.openImage(os.path.join(inputPath, sample))
#Split channels
nucleistack = cs.getChannel(imp, 1)
cellstack = cs.getChannel(imp, 2)
#Select relevant slices by average intensity of cell marker
avgI = []
for sliceIndex in range(1, cellstack.size() + 1): #loop over slices
ip = cellstack.getProcessor(sliceIndex)
avgI.append(ip.getStats().mean) #get average intensity
avgth1 = percentile(sorted(avgI), parameters['avgI_low'])
avgth2 = percentile(sorted(avgI), parameters['avgI_high'])
sel = [
avgI.index(i) + 1 for i in avgI if (i >= avgth1 and i <= avgth2)
] #pick slices with intermediate average intensity
print sel
#Get ROIs
rois = []
results = ResultsTable()
for imageFile in os.listdir(inputDir):
print "Opening " + imageFile
try:
images = BF.openImagePlus(inputDir + imageFile)
image = images[0]
except UnknownFormatException:
continue
preview = getPreview(image)
preview.show()
rm = RoiManager()
dialog = WaitForUserDialog("Action required", "Please select regions of interest in this image. Click OK when done.")
dialog.show()
rm.close()
splitter = ChannelSplitter()
imp1 = ImagePlus("CH1", splitter.getChannel(image, imageA))
imp2 = ImagePlus("CH2", splitter.getChannel(image, imageB))
title = image.getTitle()
title = title[:title.rfind('.')]
image.close()
preview.close()
ch1 = ImagePlusAdapter.wrap(imp1)
ch2 = ImagePlusAdapter.wrap(imp2)
for roi in rm.getRoisAsArray():
container = createContainer(roi, ch1, ch2)
img1 = container.getSourceImage1()
img2 = container.getSourceImage2()
mask = container.getMask()
thr1, thrimp1 = calculateThreshold(imp1, roi, methods[0])
if pixel_size is None:
if objective == 20 and microscope == "SD":
pixel_size = 0.5089
else:
pixel_size = 1.0
files = glob.glob(input_dir + "*.tif")
files.sort()
frame_i = 1
for file in files:
print("Processing frame \033[1m" + str(frame_i) + "\033[0m")
frame = IJ.openImage(file) # ImagePlus
# Get just the signal channel
frame_stack = channel_splitter.getChannel(frame, channel) # ImageStack
frame = ImagePlus("Channel " + str(channel), frame_stack)
# Map signal to entire 16-bit range
contrast_enhancer.stretchHistogram(frame, 0.01)
processor = frame.getProcessor()
if(processor.getBitDepth() == 32):
processor.setMinAndMax(0, 1.0)
else:
processor.setMinAndMax(0, processor.maxValue())
frame = ImagePlus("Frame " + str(frame_i), processor)
# Convert to 8-bit, grayscale
converter = ImageConverter(frame)
converter.convertToGray8()
