def minMax(im, min, max): # ip = im.getProcessor() # ip.setMinAndMax(min,max) # ip.applyLut() IJ.setMinAndMax(im, min, max) IJ.run(im, 'Apply LUT', '') return im
def Make8bit_UsingMinMax(_imp):
imp = _imp.duplicate()
stats = StackStatistics(imp)
IJ.setMinAndMax(imp, stats.min, stats.max)
IJ.run(imp, "8-bit", "")
IJ.setMinAndMax(imp, 0, 255)
return imp
def Objects3D(_img, impRedirect, mpar={"vmin": 2, "vmax": 100000}, _gui=False):
print "Objects3D:"
_img.show()
impRedirect.show()
options = (
"mean_gray_value centroid dots_size=5 font_size=10 show_numbers white_numbers store_results_within_a_table_named_after_the_image_(macro_friendly) redirect_to=[%s]"
% impRedirect.title
)
# print options
print "image: " + _img.title
print "redirect to: " + impRedirect.title
IJ.run("3D OC Options", options)
IJ.run(
_img,
"3D Objects Counter",
"threshold=128 slice=35 min.=%s max.=%s surfaces statistics summary" % (mpar["vmin"], mpar["vmax"]),
)
impObjects = IJ.getImage()
IJ.run(impObjects, "16 colors", "")
# somehow find the number of objects detected and set the LUT
IJ.setMinAndMax(impObjects, 0, 2)
impObjects.hide()
_img.hide()
impRedirect.hide()
return (impObjects,)
def process(self, image, seeds=None):
self.objects = None
self.labelImage = None
duplicator = Duplicator()
spots = duplicator.run(image)
IJ.setMinAndMax(spots, 0, 1)
IJ.run(spots, "16-bit", "")
spot3DImage = ImageHandler.wrap(spots)
if seeds != None:
seed3DImage = ImageHandler.wrap(seeds)
else:
seed3DImage = self.__computeSeeds(spots)
algorithm = Segment3DSpots(spot3DImage, seed3DImage)
algorithm.show = False
algorithm.setSeedsThreshold(self.seedsThreshold)
algorithm.setLocalThreshold(self.localBackground)
algorithm.setWatershed(self.watershed)
algorithm.setVolumeMin(self.volumeMin)
algorithm.setVolumeMax(self.volumeMax)
algorithm.setMethodLocal(Segment3DSpots.LOCAL_CONSTANT)
algorithm.setMethodSeg(Segment3DSpots.SEG_MAX)
algorithm.segmentAll()
self.objects = algorithm.getObjects()
self.labelImage = ImagePlus("OM_" + spots.getTitle(), algorithm.getLabelImage().getImageStack())
return self.labelImage
def modulation_contrast(raw_image_title, sir_image_title, do_map):
IJ.selectWindow(raw_image_title)
IJ.run("Modulation Contrast", "angles=3 phases=5 z_window_half-width=1")
output_images = []
if do_map:
IJ.run(
"Modulation Contrast Map", "calculate_mcnr_from_raw_data=" +
raw_image_title + " camera_bit_depth=16 or,_specify_mcnr_stack=" +
raw_image_title.rsplit('.', 1)[0] +
"_MCN reconstructed_data_stack=" + sir_image_title)
IJ.selectWindow(sir_image_title.rsplit('.', 1)[0] + '_MCM')
mcm_imp = IJ.getImage()
output_images.append(mcm_imp)
IJ.selectWindow(raw_image_title.rsplit('.', 1)[0] + '_MCN')
mcn_imp = IJ.getImage()
IJ.setMinAndMax(0, 255)
IJ.run("8-bit")
output_images.append(mcn_imp)
statistics = {}
statistics.update(parse_log('average feature MCNR = '))
statistics.update(parse_log('estimated Wiener filter optimum = '))
return output_images, statistics
def make_16bit_using_minmax(_imp): # ????? this does not make sense, or ??? imp = _imp.duplicate() stats = StackStatistics(imp) IJ.setMinAndMax(imp, stats.min, stats.max) IJ.run(imp, "16-bit", "") IJ.setMinAndMax(imp, 0, 65535) return imp
def make_16bit_using_minmax(_imp): # ????? this does not make sense, or ??? imp = _imp.duplicate() stats = StackStatistics(imp) IJ.setMinAndMax(imp, stats.min, stats.max) IJ.run(imp, "16-bit", "") IJ.setMinAndMax(imp, 0, 65535) return imp
def DistanceMap(_img, mpar={}, _gui=False):
imp = Duplicator().run(_img)
IJ.run(imp, "Exact Euclidean Distance Transform (3D)", "")
imp = IJ.getImage()
IJ.setMinAndMax(imp, 0, 65535)
IJ.run(imp, "16-bit", "")
imp.setTitle("DistanceMap")
imp.hide()
return (imp,)
def Multiply(imp1, imp2):
ic = ImageCalculator()
imp = ic.run("Multiply create stack", imp1, imp2)
stats = StackStatistics(imp)
IJ.setMinAndMax(imp, stats.min, stats.max)
print stats
imp.setTitle("Multiply")
imp.show()
return imp
def previewDisplaySettings(image, title, zoom, cal):
"""Apply wanted settings for previews"""
ImageConverter.setDoScaling(0)
ImageConverter(image).convertToGray16()
image.show()
IJ.run("glasbey_on_dark")
IJ.setMinAndMax(image, 0, 255)
image.setTitle(title)
image.setCalibration(cal)
IJ.run("Set... ", "zoom=" + str(zoom))
def extractChannel(imp, nChannel, nFrame):
"""extract a channel from the image, at a given frame returning a new imagePlus labelled with the channel name"""
stack = imp.getImageStack()
ch = ImageStack(imp.width, imp.height)
for i in range(imp.getNSlices()):
index = imp.getStackIndex(nChannel, i, nFrame)
ch.addSlice(str(i), stack.getProcessor(index))
imp3 = ImagePlus("Channel " + str(nChannel), ch).duplicate()
stats = StackStatistics(imp3)
IJ.setMinAndMax(imp3, stats.min, stats.max)
return imp3
def kotaMiuraBrightness(targetPath, fn):
'''Taken and updated from script by Kota Miura (2015) found at:
http://wiki.cmci.info/documents/120206pyip_cooking/python_imagej_cookbook#automatic_brightnesscontrast_button'''
autoThreshold = 0
AUTO_THRESHOLD = 5000
imp = IJ.getImage()
cal = imp.getCalibration()
imp.setCalibration(None)
stats = imp.getStatistics() # get uncalibrated stats
imp.setCalibration(cal)
limit = int(stats.pixelCount / 10)
histogram = stats.histogram(
) #int[] I just added the () after the histogram...?
#histogram = imp.getHistogram(256) # trying to make it bin itself in a smooth way like the java version of the code
if autoThreshold < 10:
autoThreshold = AUTO_THRESHOLD
else:
autoThreshold /= 2
threshold = int(stats.pixelCount / autoThreshold) #int
i = -1
found = False
count = 0 # int
while True:
i += 1
count = histogram[i]
if count > limit:
count = 0
found = count > threshold
if found or i >= 255:
# if 0 not in (!found, i<255) :
break
hmin = i #int
i = 256
while True:
i -= 1
count = histogram[i]
if count > limit:
count = 0
found = count > threshold
if found or i < 1:
# if 0 not in (!found, i<255) :
break
hmax = i #int
IJ.setMinAndMax(imp, hmin, hmax)
#IJ.run(imp, "8-bit", "")
IJ.run(imp, "Apply LUT", "") # this step makes histogram odd
imp.updateAndDraw()
return threshold, limit, hmin, hmax
def procAZtecImg(imp, loG, hiG, radMF, unPerPx, units=-6): """procAZtecImg(imp, loG, hiG, radMF, unPerPx, units=-6) Process an AZTec TIF image Inputs: imp - the image plus loG - the gray level to set as black hiG - the gray level to set as white radMF - the radius for a median filter unPerPx - the size of a pixel in units units - the power w.r.t. meters. Default = -6 Returns: an image plus of a duplicated image """ name = imp.getShortTitle() # make a copy wrk = imp.duplicate() IJ.run(wrk, "Median...", "radius=%g" % radMF) IJ.setMinAndMax(wrk, loG, hiG) jmg.calibImageDirect(wrk, unPerPx, units=-6) wrk.setTitle(name) return wrk
def analyse(imp, root, filename, thresholds):
closeAllImageWindows()
imp.show()
print ""
print(imp.getTitle())
print "**********************************"
IJ.run("Set Measurements...", "integrated redirect=None decimal=2");
# get image calibration info
IJ.run(imp, "Properties...", "unit=pixel pixel_width=1 pixel_height=1");
# convert to 8-bit
IJ.setMinAndMax(imp, 0, 65535);
IJ.run(imp, "8-bit", "");
thresholds = [(x / 65535 * 255) for x in thresholds]
# Nuclei
#closeAllNoneImageWindows()
iChannel = 1
imp.setSlice(iChannel)
IJ.run(imp, "Gaussian Blur...", "sigma=2 slice");
IJ.setThreshold(imp, thresholds[iChannel-1], 1000000000)
IJ.run(imp, "Convert to Mask", "method=Default background=Dark only black");
IJ.run(imp, "Measure", "");
rt = ResultsTable.getResultsTable()
values = rt.getColumnAsDoubles(rt.getColumnIndex("RawIntDen"))
print("Area Nuclei (pixel^2) =",values[-1]/255)
# Dots
#closeAllNoneImageWindows()
iChannel = 3
imp.setSlice(iChannel)
IJ.run(imp, "Gaussian Blur...", "sigma=1 slice");
IJ.run(imp, "Find Maxima...", "noise="+str(thresholds[iChannel-1])+" output=Count");
rt = ResultsTable.getResultsTable()
values = rt.getColumnAsDoubles(rt.getColumnIndex("Count"))
print("Number of Dots =",values[-1])
labId = "qm-04513"
smpId = "RAD-2015-0148Y-HCl-xs"
datDir = edsDir + "/Oxford/" + ePrjDir + "/reports/" + labId + "-" + smpId
sTifPath = datDir + "/tif/"
bSetMinMax = False
gMin = 1000
gMax = 11400
fwUnits = 28.9
units = IJ.micronSymbol + "m"
imp = IJ.getImage()
imp = jmg.calibImage(imp, fwUnits, units=-6)
if(bSetMinMax == True):
IJ.setMinAndMax(gMin, gMax)
imp.updateAndRepaintWindow()
ti = imp.getShortTitle()
fi = sTifPath + ti + ".tif"
fs = FileSaver(imp)
imp.changes = False
print(fi)
if fs.saveAsTiff(fi):
print "Tif saved successfully at ", fi
print("done")
clij2.clear()
segmentImp = extractChannel(imp1, segmentChannel, nFrame)
gfx1 = clij2.push(segmentImp)
gfx2 = clij2.create(gfx1)
gfx3 = clij2.create(gfx1)
gfx4 = clij2.create(gfx1)
gfx5 = clij2.create(gfx1)
gfx1, gfx2, gfx3, gfx4, gfx5 = segment(gfx1, gfx2, gfx3, gfx4, gfx5,
gaussianSigma, thresholdMethod,
maxIntensity, largeDoGSigma,
pixelAspect, originalTitle, topHat,
topHatSigma, manualSegment,
manualThreshold)
thresholdImp = clij2.pull(gfx3)
IJ.setMinAndMax(thresholdImp, 0, 1)
thresholdImp.setCalibration(cal)
thresholdImp.setTitle("Binary mask of " + originalTitle)
table, FRETimp2, FRETProjImp, labelImp = fretCalculations(
imp1, nFrame, donorChannel, acceptorChannel, acceptorChannel2, table,
gfx5, gfx2, gfx3, gfx4, gfx1, originalTitle)
if makeNearProj == True:
nearZImp = nearestZProject(FRETimp2)
conNearZStack = concatStacks(conNearZStack, nearZImp)
nearZImp.close()
#add the images to concatenated stacks
conThresholdStack = concatStacks(conThresholdStack, thresholdImp)
conFRETImp2Stack = concatStacks(conFRETImp2Stack, FRETimp2)
conFRETProjImpStack = concatStacks(conFRETProjImpStack, FRETProjImp)
# Apply an automatic threshold method to an image on the GPU # # Author: Robert Haase, [email protected] # April 2019 ######################################### from ij import IJ from net.haesleinhuepf.clij import CLIJ IJ.run("Close All") # load example image imp = IJ.openImage("c:/structure/code/clij-docs/src/main/resources/blobs.tif") imp.show() # init GPU clij = CLIJ.getInstance() # push image to GPU input = clij.push(imp) # reserve memory for output, same size and type as input output = clij.create(input) # apply threshold method on GPU clij.op().automaticThreshold(input, output, "Otsu") # show result clij.pull(output).show() IJ.setMinAndMax(0, 1)
def make_8bit_using_min_max(_imp): imp = _imp.duplicate() stats = StackStatistics(imp) IJ.setMinAndMax(imp, stats.min, stats.max) ImageConverter(imp).convertToGray8() return imp
gfx1 = clij2.push(segmentImp)
gfx2 = clij2.create(gfx1)
gfx3 = clij2.create(gfx1)
gfx4 = clij2.create(gfx1)
gfx5 = clij2.create(gfx1)
gfx1, gfx2, gfx3, gfx4, gfx5 = segment(gfx1, gfx2, gfx3, gfx4, gfx5,
gaussianSigma, thresholdMethod,
maxIntensity, largeDoGSigma,
pixelAspect, originalTitle, topHat,
topHatSigma, manualSegment,
manualThreshold)
thresholdImp = clij2.pull(gfx3)
labelImp = clij2.pull(gfx5)
gfx4 = clij2.push(quantImp)
IJ.setMinAndMax(thresholdImp, 0, 1)
thresholdImp.setCalibration(cal)
thresholdImp.setTitle("Binary mask of " + originalTitle)
#add the images to concatenated stacks
conThresholdStack = concatStacks(conThresholdStack, thresholdImp)
conlabelImpStack = concatStacks(conlabelImpStack, labelImp)
table = quantify(gfx4, gfx5, table, nFrame, originalTitle)
thresholdImp.close()
labelImp.close()
IJ.log("Processing timeframe: " + str(nFrame))
table.show("Results of " + originalTitle)
#Show the images and make the images pretty... I should have put in a function`
conThresholdImp = ImagePlus("Threshold image for " + originalTitle,
def analyze(iDataSet, tbModel, p):
#
# Init
#
IJ.run("Options...", "iterations=1 count=1");
close_all_image_windows()
#
# Open file
#
filepath = tbModel.getFileAbsolutePathString(iDataSet, "Input", "IMG")
if filepath.endswith('.mha'):
IJ.openImage(filepath)
imp = IJ.getImage()
# get rid of the unsigned int problem
IJ.run(imp, "32-bit", "");
IJ.setMinAndMax(0, 65535);
IJ.run(imp, "16-bit", "");
elif filepath.endswith('.h5'):
imp = None
IJ.run(imp, "Scriptable load HDF5...", "load="+filepath+" datasetnames="+p["HDF5_data_set_name"]+" nframes=1 nchannels=1");
imp = IJ.getImage()
else:
imp = IJ.openImage(filepath)
imp.show()
#
# Crop after loading
#
if p['crop']:
x_min = int(p['mask_roi'][0])
y_min = int(p['mask_roi'][1])
z_min = int(p['mask_roi'][2])
x_width = int(p['mask_roi'][3])
y_width = int(p['mask_roi'][4])
z_width = int(p['mask_roi'][5])
stack = imp.getImageStack()
stack_cropped = stack.crop(x_min, y_min, z_min, x_width, y_width, z_width)
imp = ImagePlus('',stack)
#
# Convert
#
IJ.setMinAndMax(p['map_to_zero'], p['map_to_max']);
IJ.run(imp, p['output_bit_depth'], "");
#
# Scale (Binning)
#
if p['binning']:
sx = float(1.0/p['binning_x'])
sy = float(1.0/p['binning_y'])
sz = float(1.0/p['binning_z'])
parameters = "x="+str(sx)+" y="+str(sy)+" z="+str(sz)+" interpolation=Bilinear average process create"
IJ.run(imp, "Scale...", parameters);
imp = IJ.getImage()
#
# Save converted volume data
#
if p['save_volume_data']:
converted_filename = tbModel.getFileName(iDataSet, "Input", "IMG")+"--converted."+p['output_format']
tbModel.setFileAbsolutePath(p['output_folder'], converted_filename, iDataSet, 'Output', 'IMG')
IJ.saveAs(imp, p['output_format'], tbModel.getFileAbsolutePathString(iDataSet, 'Output', 'IMG'))
#
# Projections
#
if p['save_xyz_projections']:
# compute one XYZ projection in one image
projection = 'XYZ'
imp_projected = project_XYZ(imp)
# construct output filename
projected_filename = tbModel.getFileName(iDataSet, "Input", "IMG")+"--"+projection+"."+p['output_format']
# store in table
tbModel.setFileAbsolutePath(p['output_folder'], projected_filename, iDataSet, projection, "IMG")
# store on disk
IJ.saveAs(imp_projected,p['output_format'],tbModel.getFileAbsolutePathString(iDataSet, projection, "IMG"))
'''
projections = ['X','Y','Z']
for projection in projections:
# compute projection
imp_projected = project(imp, projection)
# construct output filename
projected_filename = tbModel.getFileName(iDataSet, "Input", "IMG")+"--"+projection+"."+p['output_format']
# store in table
tbModel.setFileAbsolutePath(p['output_folder'], projected_filename, iDataSet, projection, "IMG")
# store on disk
IJ.saveAs(imp_projected,p['output_format'],tbModel.getFileAbsolutePathString(iDataSet, projection, "IMG"))
'''
return tbModel
tracked[i]= 11
print test.labelValues
fp= ShortProcessor(len(tracked), 1,tracked , None)
labelerImp= ImagePlus("labeler", fp)
src2=clij2.push(labelerImp)
conLabeledStack=ImageStack(imp1.width, imp1.height)
if frames>1:
for nFrame in range(1,frames+1):
imp3=extractFrame(imp1, nFrame)
src=clij2.push(imp3)
dst=clij2.create(src)
clij2.replaceIntensities(src, src2, dst)
LabeledImp=clij2.pull(dst)
conLabeledStack = concatStacks( conLabeledStack, LabeledImp)
concatLabeledImp= ImagePlus("Labeled "+imageName, conLabeledStack)
ImageConverter.setDoScaling(0)
ImageConverter(concatLabeledImp).convertToGray16()
IJ.setMinAndMax(concatLabeledImp, 0, 255)
concatLabeledImp.setCalibration(imp1.getCalibration())
concatLabeledImp.setDimensions(1, imp1.getNSlices(), imp1.getNFrames())
concatLabeledImp = CompositeImage(concatLabeledImp, CompositeImage.COMPOSITE)
concatLabeledImp.show()
IJ.run("glasbey_on_dark")
labelerImp.close()
def convert_to_8bit(imp, vmin, vmax): IJ.setMinAndMax(imp, vmin, vmax) ImageConverter(imp).convertToGray8() return imp
def previewDialog(imp):
gd = GenericDialogPlus("Nuclear segmentation and quantification v1.01")
#create a list of the channels in the provided imagePlus
types = []
for i in xrange(1, imp.getNChannels() + 1):
types.append(str(i))
#user can pick which channel to base the segmentation on
gd.addChoice("Channel number to use for segmentation", types, types[0])
gd.addChoice("Channel to quantify", types, types[0])
methods = [
"Otsu", "Default", "Huang", "Intermodes", "IsoData", "IJ_IsoData",
"Li", "MaxEntropy", "Mean", "MinError", "Minimum", "Moments",
"Percentile", "RenyiEntropy", "Shanbhag", "Triangle", "Yen"
]
gd.addChoice("Autosegmentation method", methods, methods[0])
intensities = ["254", "4094", "65534"]
gd.addChoice("Max Intensity", intensities, intensities[-1])
gd.addSlider("Small DoG sigma", 0.5, 10, 1, 0.1)
gd.addSlider("Large DoG sigma", 0.5, 20, 5, 0.1)
gd.addCheckbox("TopHat background subtraction? (Slower, but better) ",
True)
gd.addSlider("TopHat sigma", 5, 20, 8, 0.1)
gd.setModal(False)
gd.addCheckbox("Manually set threshold? ", False)
gd.addSlider("Manual threshold", 10, 65534, 2000, 1)
gd.hideCancelButton()
gd.showDialog()
cal = imp.getCalibration()
pixelAspect = (cal.pixelDepth / cal.pixelWidth)
originalTitle = imp1.getTitle()
choices = gd.getChoices()
print choices
sliders = gd.getSliders()
checkboxes = gd.getCheckboxes()
segmentChannel = int(choices.get(0).getSelectedItem())
quantChannel = int(choices.get(1).getSelectedItem())
thresholdMethod = choices.get(2).getSelectedItem()
maxIntensity = int(choices.get(3).getSelectedItem())
gaussianSigma = sliders.get(0).getValue() / 10.0
largeDoGSigma = gd.sliders.get(1).getValue() / 10.0
topHat = gd.checkboxes.get(0).getState()
topHatSigma = gd.sliders.get(2).getValue() / 10.0
manualSegment = gd.checkboxes.get(1).getState()
manualThreshold = gd.sliders.get(3).getValue()
segmentChannelOld = segmentChannel
thresholdMethodOld = thresholdMethod
maxIntensityOld = maxIntensity
gaussianSigmaOld = gaussianSigma
largeDoGSigmaOld = largeDoGSigma
topHatOld = topHat
topHatSigmaOld = topHatSigma
manualSegmentOld = manualSegment
manualThresholdOld = manualThreshold
clij2.clear()
segmentImp = extractChannel(imp1, segmentChannel, 0)
try:
gfx1 = clij2.push(segmentImp)
gfx2 = clij2.create(gfx1)
gfx3 = clij2.create(gfx1)
gfx4 = clij2.create(gfx1)
gfx5 = clij2.create(gfx1)
gfx7 = clij2.create([imp.getWidth(), imp.getHeight()])
except:
try:
Thread.sleep(500)
IJ.log(
"Succeeded to sending to graphics card on the second time...")
gfx1 = clij2.push(segmentImp)
gfx2 = clij2.create(gfx1)
gfx3 = clij2.create(gfx1)
gfx4 = clij2.create(gfx1)
gfx5 = clij2.create(gfx1)
gfx7 = clij2.create([imp.getWidth(), imp.getHeight()])
except:
errorDialog(
"""Could not send image to graphics card, it may be too large!
Easy solutions: Try processing as 8-bit, cropping or scaling the image, or
select a different CLIJ2 GPU.
This issue is often intermittent, so trying again may also work!
See the "Big Images on x graphics cards' notes at:
https://clij2.github.io/clij2-docs/troubleshooting for more solutions
""" + str(clij2.reportMemory()))
gfx1, gfx2, gfx3, gfx4, gfx5 = segment(gfx1, gfx2, gfx3, gfx4, gfx5,
gaussianSigma, thresholdMethod,
maxIntensity, largeDoGSigma,
pixelAspect, originalTitle, topHat,
topHatSigma, manualSegment,
manualThreshold)
clij2.maximumZProjection(gfx5, gfx7)
labelPrevImp = clij2.pull(gfx7)
IJ.setMinAndMax(labelPrevImp, 0, clij2.getMaximumOfAllPixels(gfx7))
labelPrevImp.setTitle("Preview segmentation")
labelPrevImp.show()
IJ.run("glasbey_inverted")
while ((not gd.wasCanceled()) and not (gd.wasOKed())):
segmentChannel = int(choices.get(0).getSelectedItem())
quantChannel = int(choices.get(1).getSelectedItem())
thresholdMethod = choices.get(2).getSelectedItem()
maxIntensity = int(choices.get(3).getSelectedItem())
gaussianSigma = sliders.get(0).getValue() / 10.0
largeDoGSigma = gd.sliders.get(1).getValue() / 10.0
topHat = gd.checkboxes.get(0).getState()
topHatSigma = gd.sliders.get(2).getValue() / 10.0
manualSegment = gd.checkboxes.get(1).getState()
manualThreshold = gd.sliders.get(3).getValue()
if (segmentChannelOld != segmentChannel
or thresholdMethodOld != thresholdMethod
or maxIntensityOld != maxIntensity
or gaussianSigmaOld != gaussianSigma
or largeDoGSigmaOld != largeDoGSigma or topHatOld != topHat
or topHatSigmaOld != topHatSigma
or manualSegmentOld != manualSegment
or manualThresholdOld != manualThreshold):
if segmentChannelOld != segmentChannel:
clij2.clear()
segmentImp = extractChannel(imp1, segmentChannel, 0)
gfx1 = clij2.push(segmentImp)
gfx2 = clij2.create(gfx1)
gfx3 = clij2.create(gfx1)
gfx4 = clij2.create(gfx1)
gfx5 = clij2.create(gfx1)
gfx7 = clij2.create([imp.getWidth(), imp.getHeight()])
gfx1, gfx2, gfx3, gfx4, gfx5 = segment(
gfx1, gfx2, gfx3, gfx4, gfx5, gaussianSigma, thresholdMethod,
maxIntensity, largeDoGSigma, pixelAspect, originalTitle,
topHat, topHatSigma, manualSegment, manualThreshold)
clij2.maximumZProjection(gfx5, gfx7)
labelPrevImp.close()
labelPrevImp = clij2.pull(gfx7)
IJ.setMinAndMax(labelPrevImp, 0, clij2.getMaximumOfAllPixels(gfx7))
labelPrevImp.setTitle("Preview segmentation")
labelPrevImp.show()
IJ.run("glasbey_inverted")
segmentChannelOld = segmentChannel
thresholdMethodOld = thresholdMethod
maxIntensityOld = maxIntensity
gaussianSigmaOld = gaussianSigma
largeDoGSigmaOld = largeDoGSigma
topHatOld = topHat
topHatSigmaOld = topHatSigma
manualSegmentOld = manualSegment
manualThresholdOld = manualThreshold
Thread.sleep(200)
labelPrevImp.close()
return segmentChannel, quantChannel, thresholdMethod, maxIntensity, gaussianSigma, largeDoGSigma, topHat, topHatSigma, manualSegment, manualThreshold
def run():
### Default arguments
two_sarcomere_size = 25 # determined by filter used.
rotation_angle = 0.0
### Get the image we'd like to work with.
# Don't need to ask where to save the intermediate image since it'll just be saved in the matchedmyo folder anyway
# may change this though. In case they want to keep that image around.
this_img = WindowManager.getCurrentImage()
if this_img == None:
ud = WaitForUserDialog(
"Please select the image you would like to analyze.")
ud.show()
this_img = WindowManager.getCurrentImage()
img_name = this_img.getTitle()
matchedmyo_path = "/home/AD/dfco222/scratchMarx/matchedmyo/" # this would be grabbed from the prompt
gd = GenericDialog("Preprocessing Options")
gd.addCheckbox("Automatic Resizing/Rotation", True)
gd.addCheckbox("CLAHE", True)
gd.addCheckbox("Normalization to Transverse Tubules", True)
gd.showDialog()
if gd.wasCanceled():
return
auto_resize_rotate = gd.getNextBoolean()
clahe = gd.getNextBoolean()
normalize = gd.getNextBoolean()
if auto_resize_rotate:
# Clear selection so it takes FFT of full image
rotation_angle = auto_resize_angle_measure(this_img,
two_sarcomere_size)
if clahe:
clahe_args = "blocksize={} histogram=256 maximum=3 mask=*None* fast_(less_accurate)".format(
two_sarcomere_size)
IJ.run(this_img, "Enhance Local Contrast (CLAHE)", clahe_args)
if normalize:
# Ask the user to select a subsection of the image that looks healthy-ish.
ud = WaitForUserDialog(
"Please select a subsection exhibiting a measure of healthy TT structure using the Rectangle tool.\n"
+ " Only a single cell or several are needed.\n\n" +
" Press 'OK' when finished.")
ud.show()
IJ.setTool("rectangle")
# Duplicate the selected subsection.
selection = this_img.crop()
IJ.run(selection, "Duplicate...", "title=subsection.tif")
# Grab the subsection image and rotate it.
selection = WindowManager.getImage("subsection.tif")
IJ.run(
selection, "Rotate...",
"angle={} grid=1 interpolation=Bicubic enlarge".format(
rotation_angle))
# Ask the user to select a bounding box that contains only tubules
# NOTE: Need to get rid of initial selection since it's the entire image and it's annoying to click out of
IJ.setTool("rectangle")
IJ.run(selection, "Select None", "")
ud = WaitForUserDialog(
"Select a subsection of the image that contains only tubules and no membrane."
)
ud.show()
# Grab the subsection ImagePlus
selection = WindowManager.getCurrentImage()
this_window = WindowManager.getActiveWindow()
selection_small = selection.crop()
IJ.run(selection, "Close", "")
# NOTE: May not actually display this depending on how the macros work
IJ.run(selection_small, "Duplicate...", "title=subsection_small.tif")
# Smooth the selection using the single TT filter.
# NOTE: It won't read in so we're just going to hard code it in since it's simple
tt_filt_row = "0 0 0 1 1 1 1 1 1 0 0 0 0\n"
tt_filt = ""
for i in range(21):
tt_filt += tt_filt_row
IJ.run("Convolve...", "text1=[" + tt_filt + "] normalize")
# Segment out the TTs from the 'gaps' using Gaussian Adaptive Thresholding.
selection_small = WindowManager.getImage("subsection_small.tif")
IJ.run(selection_small, "Duplicate...", "title=thresholded.tif")
threshed = WindowManager.getImage("thresholded.tif")
IJ.run(threshed, "Auto Local Threshold",
"method=Bernsen radius=7 parameter_1=1 parameter_2=0 white")
# Select the TTs from the thresholded image.
IJ.run(threshed, "Create Selection", "")
tt_selection = WindowManager.getImage("subsection_small.tif")
IJ.selectWindow("thresholded.tif")
IJ.selectWindow("subsection_small.tif")
IJ.run(tt_selection, "Restore Selection", "")
# Get TT intensity statistics.
stat_options = IS.MEAN | IS.MIN_MAX | IS.STD_DEV
stats = IS.getStatistics(tt_selection.getProcessor(), stat_options,
selection_small.getCalibration())
# Calculate pixel ceiling intensity value based on heuristic.
# TODO: Add a catch for data type overflow.
pixel_ceiling = stats.mean + 3 * stats.stdDev
print "px ceil:", pixel_ceiling
# Invert selection to get inter-sarcomeric gap intensity statistics.
IJ.run(tt_selection, "Make Inverse", "")
stat_options = IS.MEAN | IS.MIN_MAX | IS.STD_DEV
stats = IS.getStatistics(tt_selection.getProcessor(), stat_options,
selection_small.getCalibration())
# Calculate pixel floor intensity value based on heuristic.
pixel_floor = stats.mean - stats.stdDev
# TODO: Add a catch for data type underflow.
print "px floor:", pixel_floor
# Threshold original image based on these values.
IJ.selectWindow(this_img.getTitle())
IJ.run(this_img, "Select All", "")
IJ.setMinAndMax(pixel_floor, pixel_ceiling)
IJ.run(this_img, "Apply LUT", "")
## Ask if it is acceptable.
gd = GenericDialog("Acceptable?")
gd.addMessage(
"If the preprocessed image is acceptable for analysis, hit 'OK' to being analysis.\n"
+ " If the image is unacceptable or an error occurred, hit 'Cancel'")
gd.showDialog()
if gd.wasCanceled():
return
## Save the preprocessed image.
imp = IJ.getImage()
fs = FileSaver(imp)
img_save_dir = matchedmyo_path + "myoimages/" # actually get from user at some point
img_file_path = img_save_dir + img_name[:-4] + "_preprocessed.tif"
if os.path.exists(img_save_dir) and os.path.isdir(img_save_dir):
print "Saving image as:", img_file_path
if os.path.exists(img_file_path):
# use dialog box to ask if they want to overwrite
gd = GenericDialog("Overwrite?")
gd.addMessage(
"A file exists with the specified path, \"{}\". Would you like to overwrite it?"
.format(img_file_path))
gd.enableYesNoCancel()
gd.showDialog()
if gd.wasCanceled():
return
elif fs.saveAsTiff(img_file_path):
print "Preprocessed image saved successfully at:", '"' + img_file_path + '"'
else:
print "Folder does not exist or is not a folder!"
### Create the YAML file containing the parameters for classification
## Ask user for YAML input
gd = GenericDialog("YAML Input")
gd.addStringField("imageName", img_file_path, 50)
#gd.addStringField("maskName", "None")
gd.addStringField("outputParams_fileRoot", img_file_path[:-4], 50)
gd.addStringField("outputParams_fileType", "tif")
gd.addNumericField("outputParams_dpi", 300, 0)
gd.addCheckbox("outputParams_saveHitsArray", False)
gd.addStringField("outputParams_csvFile", matchedmyo_path + "results/")
gd.addCheckbox("TT Filtering", True)
gd.addToSameRow()
gd.addCheckbox("LT Filtering", True)
gd.addToSameRow()
gd.addCheckbox("TA Filtering", True)
gd.addNumericField("scopeResolutions_x", 5.0, 3)
gd.addToSameRow()
gd.addNumericField("scopeResolutions_y", 5.0, 3)
gd.addMessage("Enter in filter rotation angles separated by commas.")
gd.addStringField("", "-25, -20, -15, -10, -5, 0, 5, 10, 15, 20, 25", 50)
gd.addCheckbox("returnAngles", False)
gd.addCheckbox("returnPastedFilter", True)
gd.showDialog()
if gd.wasCanceled():
return
strings = [st.text for st in gd.getStringFields()]
#if strings[1] == "None" or "":
# strings[1] = None
nums = [float(num.text) for num in gd.getNumericFields()]
nums[0] = int(nums[0]) # Have to make sure the dpi variable is an integer
checks = [str(bool(boo.state)) for boo in gd.getCheckboxes()]
iter_argument = ','.join(
[str(float(it) - rotation_angle) for it in strings[4].split(',')])
string_block = """imageName: {0[0]}
outputParams:
fileRoot: {0[1]}
fileType: {0[2]}
dpi: {1[0]}
saveHitsArray: {2[0]}
csvFile: {0[3]}
preprocess: False
filterTypes:
TT: {2[1]}
LT: {2[2]}
TA: {2[3]}
scopeResolutions:
x: {1[1]}
y: {1[2]}
iters: [{3}]
returnAngles: {2[4]}
returnPastedFilter: {2[5]}""".format(strings, nums, checks, iter_argument)
im_title = this_img.getTitle()
with cd(matchedmyo_path):
yaml_file_path = "./YAML_files/" + im_title[:-4] + ".yml"
with open("./YAML_files/" + im_title[:-4] + ".yml", "w") as ym:
ym.write(string_block)
print "Wrote YAML file to:", matchedmyo_path + yaml_file_path[2:]
### Run the matchedmyo code on the preprocessed image
with cd(matchedmyo_path):
#os.chdir(matchedmyo_path)
#subprocess.call(["python3", matchedmyo_path+"matchedmyo.py", "fullValidation"])
subprocess.call(
["python3", "matchedmyo.py", "run", "--yamlFile", yaml_file_path])
def convert_to_8bit(imp, vmin, vmax): IJ.setMinAndMax(imp, vmin, vmax) ImageConverter(imp).convertToGray8() return imp
for i in range(0, 36): # change the shift from slice to slice at = AffineTransform2D(); at.translate(-input.getWidth() / 2, -input.getHeight() / 2); at.rotate(i * 10.0 / 180.0 * Math.PI); at.translate(input.getWidth() / 2, input.getHeight() / 2); clij.op().affineTransform2D(shiftXgpu, rotatedShiftXgpu, at); # apply transform clij.op().applyVectorfield(input, rotatedShiftXgpu, shiftYgpu, temp); # put resulting 2D image in the right plane clij.op().copySlice(temp, output, i); # show result clij.pull(output).show(); IJ.setMinAndMax(0, 255); input.close(); shiftXgpu.close(); rotatedShiftXgpu.close(); shiftYgpu.close(); temp.close(); output.close();
def processImage(filename):
# many zeiss images have a pyramid format. In order to speed up the evaluation, we chose only the second highest resolution (=series_2).
# depending on your computer/workstation this parameter can be optimized (i.e. chose series_1 if you have a fast computer or series_3 for slow ones)
IJ.run(
"Bio-Formats", "open=" + filename +
" color_mode=Composite rois_import=[ROI manager] view=Hyperstack stack_order=XYCZT series_2"
)
imp = IJ.getImage()
imp.show()
if (not entertain):
imp.show()
# preparing memory for saving the results
original1 = NewImage.createFloatImage("Original 1",
imp.getWidth() / tileSize + 1,
imp.getHeight() / tileSize + 1, 1,
NewImage.FILL_BLACK)
original1Map = original1.getProcessor()
original2 = NewImage.createFloatImage("Original 2",
imp.getWidth() / tileSize + 1,
imp.getHeight() / tileSize + 1, 1,
NewImage.FILL_BLACK)
original2Map = original2.getProcessor()
spotCount1 = NewImage.createFloatImage("Spot count 1",
imp.getWidth() / tileSize + 1,
imp.getHeight() / tileSize + 1, 1,
NewImage.FILL_BLACK)
spotCount1Map = spotCount1.getProcessor()
spotCount2 = NewImage.createFloatImage("Spot count 2",
imp.getWidth() / tileSize + 1,
imp.getHeight() / tileSize + 1, 1,
NewImage.FILL_BLACK)
spotCount2Map = spotCount2.getProcessor()
ratio = NewImage.createFloatImage("Ratio",
imp.getWidth() / tileSize + 1,
imp.getHeight() / tileSize + 1, 1,
NewImage.FILL_BLACK)
ratioMap = ratio.getProcessor()
if (entertain):
ratio.show()
# go through all tiles
for x in range(0, ratioMap.getWidth() - 1):
for y in range(0, ratioMap.getHeight() - 1):
# crop out the tile from the original images
imp.setRoi(Roi(x * tileSize, y * tileSize, tileSize, tileSize))
channel1 = Duplicator().run(imp, 1, 1, 1, 1, 1, 1)
channel2 = Duplicator().run(imp, 2, 2, 1, 1, 1, 1)
# spot detection
spots1 = detectSpots(channel1,
spotDetection_GaussianBlur_sigma_GFP,
spotDetection_findMaxima_noise_GFP)
spots2 = detectSpots(channel2,
spotDetection_GaussianBlur_sigma_DAPI,
spotDetection_findMaxima_noise_DAPI)
# pixel statistics
statistics1 = channel1.getStatistics()
statistics2 = channel2.getStatistics()
# calculate ratio if spots were found
s1 = 0
s2 = 0
r = 0
if (spots1 is not None and spots2 is not None):
fp1 = spots1.getFloatPolygon()
fp2 = spots2.getFloatPolygon()
s1 = fp1.npoints
s2 = fp2.npoints
if (s2 > cutOff_DAPI and s1 > cutOff_GFP):
r = 1.0 * s1 / s2
# fill result memory
original1Map.setf(x, y, statistics1.mean)
original2Map.setf(x, y, statistics2.mean)
spotCount1Map.setf(x, y, s1)
spotCount2Map.setf(x, y, s2)
ratioMap.setf(x, y, r)
if (entertain):
# show current result image
ratio.updateAndDraw()
IJ.run(ratio, "Enhance Contrast", "saturated=0.35")
# put all results image channels together to one image
images = []
images.append(original1)
images.append(original2)
images.append(spotCount1)
images.append(spotCount2)
images.append(ratio)
resultMap = RGBStackMerge.mergeChannels(images, False)
# fix pixel size
# factor is multiplied by 2 because ImageJ seems to have a problem when using .czi file series of lower resolution (i.e. series_2); please check for individual cases!
factor = (imp.getWidth() / resultMap.getWidth()) * 2
IJ.run(
resultMap, "Properties...", "channels=5 slices=1 frames=1 unit=" +
imp.getCalibration().getUnit() + " pixel_width=" +
str(imp.getCalibration().pixelWidth * factor) + " pixel_height=" +
str(imp.getCalibration().pixelHeight * factor) + " voxel_depth=1.0000")
IJ.run(
ratio, "Properties...", "channels=1 slices=1 frames=1 unit=" +
imp.getCalibration().getUnit() + " pixel_width=" +
str(imp.getCalibration().pixelWidth * factor) + " pixel_height=" +
str(imp.getCalibration().pixelHeight * factor) + " voxel_depth=1.0000")
# visualisation
resultMap.setC(1)
IJ.run(resultMap, "Green", "")
IJ.run(resultMap, "Enhance Contrast", "saturated=0.35")
resultMap.setC(2)
IJ.run(resultMap, "Blue", "")
IJ.run(resultMap, "Enhance Contrast", "saturated=0.35")
resultMap.setC(3)
IJ.run(resultMap, "mpl-inferno", "")
IJ.run(resultMap, "Enhance Contrast", "saturated=0.35")
resultMap.setC(4)
IJ.run(resultMap, "mpl-inferno", "")
IJ.run(resultMap, "Enhance Contrast", "saturated=0.35")
resultMap.setC(5)
IJ.run(resultMap, "mpl-inferno", "")
resultMap.show()
IJ.resetMinAndMax(resultMap)
resultMap.setDisplayMode(IJ.COLOR)
IJ.run(ratio, "mpl-inferno", "")
IJ.setMinAndMax(ratio, 0, 1)
# save result
IJ.saveAsTiff(resultMap, filename + "_suitable-name_map.tif")
IJ.saveAsTiff(ratio, filename + "_suitable-name_ratio.tif")
IJ.run("Close All", "")
# simpleSetHiLoDisplayLimits.py
#
# Manually set the Hi and Low display limits for a 16 or 32 bit image
#
# Modifications
# Date Who Ver What
# ---------- --- ------ -------------------------------------------------
# 2017-10-1p JRM 0.0.01 prototype
from org.python.core import codecs
codecs.setDefaultEncoding('utf-8')
import os
import glob
import time
from ij import IJ, Prefs
gLo = 1800
gHi = 14973
orig = IJ.getImage()
bd = orig.getBitDepth()
if(bd==16):
IJ.setMinAndMax(orig, gLo, gHi)
elif(bd==32):
IJ.setMinAndMax(orig, gLo, gHi)
def previewDialog(imp):
gd = GenericDialogPlus("FRETENATOR")
#create a list of the channels in the provided imagePlus
types = []
for i in xrange(1, imp.getNChannels()+1):
types.append(str(i))
gd.addMessage("""Rowe, J. H, Rizza, A., Jones A. M. (2022) Quantifying phytohormones
in vivo with FRET biosensors and the FRETENATOR analysis toolset
Methods in Molecular Biology
Rowe, JH., Grangé-Guermente, M., Exposito-Rodriguez, M.,Wimalasekera, R., Lenz, M.,
Shetty, K., Cutler, S., Jones, AM., Next-generation ABACUS biosensors reveal cellular
ABA dynamics driving root growth at low aerial humidity
""")
#user can pick which channel to base the segmentation on
if len(types)>2:
gd.addChoice("Channel number to use for segmentation", types, types[2])
gd.addChoice("Channel number to use for donor", types, types[0])
gd.addChoice("Channel number to use for acceptor (FRET)", types, types[1])
gd.addChoice("Channel number to use for acceptor", types, types[2])
#print('YAY')
else:
gd.addChoice("Channel number to use for segmentation", types, types[-1])
gd.addChoice("Channel number to use for donor", types, types[0])
gd.addChoice("Channel number to use for acceptor (FRET)", types, types[-2])
gd.addChoice("Channel number to use for acceptor", types, types[-1])
methods=["Otsu","Default", "Huang", "Intermodes", "IsoData", "IJ_IsoData", "Li", "MaxEntropy", "Mean", "MinError", "Minimum", "Moments", "Percentile", "RenyiEntropy", "Shanbhag", "Triangle", "Yen"]
gd.addChoice("Autosegmentation method", methods, methods[0])
intensities=["254", "4094", "65534"]
gd.addChoice("Max Intensity", intensities, intensities[-1])
gd.addSlider("Small DoG sigma", 0.5, 10, 0.8, 0.1)
gd.addSlider("Large DoG sigma", 0.5, 20, 4 ,0.1)
gd.addCheckbox("TopHat background subtraction? (Slower, but better) ", False)
gd.addSlider("TopHat sigma", 5, 20, 8 ,0.1)
gd.setModal(False)
gd.addCheckbox("Manually set threshold? ", False)
gd.addSlider("Manual threshold", 10, 65534, 2000, 1)
dilationOptions=["0", "1", "2","3", "4", "5", "6"]
gd.addChoice("Dilation?", dilationOptions, "0")
gd.addCheckbox("Size exclusion of ROI? ", False)
gd.addSlider("Minimum ROI size", 0, 9999, 20, 1)
gd.addSlider("Maximum ROI size", 1, 10000, 10000, 1)
gd.addCheckbox("Create nearest point projection with outlines (SLOW)? ", True)
gd.addCheckbox("Watershed object splitting? ", True)
gd.showDialog()
cal = imp.getCalibration()
pixelAspect=(cal.pixelDepth/cal.pixelWidth)
originalTitle=imp1.getTitle()
choices=gd.getChoices()
sliders=gd.getSliders()
checkboxes=gd.getCheckboxes()
segmentChannel=int(choices.get(0).getSelectedItem())
donorChannel=int(choices.get(1).getSelectedItem())
acceptorChannel=int(choices.get(2).getSelectedItem())
acceptorChannel2=int(choices.get(3).getSelectedItem())
thresholdMethod=choices.get(4).getSelectedItem()
maxIntensity=int(choices.get(5).getSelectedItem())
gaussianSigma=sliders.get(0).getValue()/10.0
largeDoGSigma = gd.sliders.get(1).getValue()/10.0
topHat=gd.checkboxes.get(0).getState()
topHatSigma=gd.sliders.get(2).getValue()/10.0
manualSegment = gd.checkboxes.get(1).getState()
manualThreshold=gd.sliders.get(3).getValue()
dilation=int(choices.get(6).getSelectedItem())
sizeExclude=gd.checkboxes.get(2).getState()
minSize = gd.sliders.get(4).getValue()
maxSize = gd.sliders.get(5).getValue()
watershed = gd.checkboxes.get(4).getState()
#print dir(gd.sliders.get(5))
#print maxSize
segmentChannelOld=segmentChannel
thresholdMethodOld=thresholdMethod
maxIntensityOld=maxIntensity
gaussianSigmaOld=gaussianSigma
largeDoGSigmaOld= largeDoGSigma
topHatOld=topHat
topHatSigmaOld=topHatSigma
manualSegmentOld= manualSegment
manualThresholdOld=manualThreshold
dilationOld=dilation
sizeExcludeOld=sizeExclude
minSizeOld=minSize
maxSizeOld=maxSize
watershedOld=watershed
clij2.clear()
segmentImp=extractChannel(imp1, segmentChannel, 0)
try:
gfx1=clij2.push(segmentImp)
gfx2=clij2.create(gfx1)
gfx3=clij2.create(gfx1)
gfx4=clij2.create(gfx1)
gfx5=clij2.create(gfx1)
gfx7=clij2.create([imp.getWidth(), imp.getHeight()])
except:
try:
Thread.sleep(500)
print("Succeeded to sending to graphics card on the second time...")
gfx1=clij2.push(segmentImp)
gfx2=clij2.create(gfx1)
gfx3=clij2.create(gfx1)
gfx4=clij2.create(gfx1)
gfx5=clij2.create(gfx1)
gfx7=clij2.create([imp.getWidth(), imp.getHeight()])
except:
errorDialog("""Could not send image to graphics card, it may be too large!
Easy solutions: Try processing as 8-bit, cropping or scaling the image, or
select a different CLIJ2 GPU.
This issue is often intermittent, so trying again may also work!
See the "Big Images on x graphics cards' notes at:
https://clij2.github.io/clij2-docs/troubleshooting for more solutions
""" + str(clij2.reportMemory()) )
gfx1,gfx2,gfx3,gfx4,gfx5 = segment(gfx1,gfx2,gfx3,gfx4,gfx5, gaussianSigma, thresholdMethod,maxIntensity, largeDoGSigma, pixelAspect, originalTitle, topHat, topHatSigma , manualSegment, manualThreshold, dilation,sizeExclude, minSize, maxSize, watershed)
clij2.maximumZProjection(gfx5, gfx7)
labelPrevImp= clij2.pull(gfx7)
IJ.setMinAndMax(labelPrevImp, 0,clij2.getMaximumOfAllPixels(gfx7))
labelPrevImp.setTitle("Preview segmentation")
labelPrevImp.show()
IJ.run("glasbey_inverted")
while ((not gd.wasCanceled()) and not (gd.wasOKed())):
segmentChannel=int(choices.get(0).getSelectedItem())
donorChannel=int(choices.get(1).getSelectedItem())
acceptorChannel=int(choices.get(2).getSelectedItem())
acceptorChannel2=int(choices.get(3).getSelectedItem())
thresholdMethod=choices.get(4).getSelectedItem()
maxIntensity=int(choices.get(5).getSelectedItem())
gaussianSigma=sliders.get(0).getValue()/10.0
largeDoGSigma = gd.sliders.get(1).getValue()/10.0
topHat=gd.checkboxes.get(0).getState()
topHatSigma=gd.sliders.get(2).getValue()/10.0
manualSegment = gd.checkboxes.get(1).getState()
manualThreshold = gd.sliders.get(3).getValue()
dilation=int(choices.get(6).getSelectedItem())
sizeExclude=gd.checkboxes.get(2).getState()
minSize = gd.sliders.get(4).getValue()
maxSize = gd.sliders.get(5).getValue()
watershed = gd.checkboxes.get(4).getState()
if (segmentChannelOld !=segmentChannel or
thresholdMethodOld !=thresholdMethod or
maxIntensityOld !=maxIntensity or
gaussianSigmaOld !=gaussianSigma or
largeDoGSigmaOld != largeDoGSigma or
topHatOld !=topHat or
topHatSigmaOld !=topHatSigma or
manualSegmentOld != manualSegment or
manualThresholdOld !=manualThreshold or
dilation != dilationOld or
sizeExcludeOld!=sizeExclude or
minSizeOld!=minSize or
maxSizeOld!=maxSize or
watershedOld!=watershed
):
if minSizeOld!=minSize:
if minSize>=maxSize:
maxSize=minSize+1
gd.sliders.get(5).setValue(maxSize)
if maxSizeOld!=maxSize:
if minSize>=maxSize:
minSize=maxSize-1
gd.sliders.get(4).setValue(minSize)
if segmentChannelOld!=segmentChannel:
clij2.clear()
segmentImp=extractChannel(imp1, segmentChannel, 0)
gfx1=clij2.push(segmentImp)
gfx2=clij2.create(gfx1)
gfx3=clij2.create(gfx1)
gfx4=clij2.create(gfx1)
gfx5=clij2.create(gfx1)
gfx7=clij2.create([imp.getWidth(), imp.getHeight()])
gfx1,gfx2,gfx3,gfx4,gfx5 = segment(gfx1,gfx2,gfx3,gfx4,gfx5, gaussianSigma, thresholdMethod,maxIntensity, largeDoGSigma, pixelAspect, originalTitle, topHat,topHatSigma, manualSegment, manualThreshold, dilation,sizeExclude, minSize, maxSize, watershed)
clij2.maximumZProjection(gfx5, gfx7)
labelPrevImp.close()
labelPrevImp= clij2.pull(gfx7)
IJ.setMinAndMax(labelPrevImp, 0,clij2.getMaximumOfAllPixels(gfx7))
labelPrevImp.setTitle("Preview segmentation")
labelPrevImp.show()
IJ.run("glasbey_inverted")
segmentChannelOld=segmentChannel
thresholdMethodOld=thresholdMethod
maxIntensityOld=maxIntensity
gaussianSigmaOld=gaussianSigma
largeDoGSigmaOld = largeDoGSigma
topHatOld=topHat
topHatSigmaOld=topHatSigma
manualSegmentOld= manualSegment
manualThresholdOld=manualThreshold
dilationOld=dilation
sizeExcludeOld=sizeExclude
minSizeOld=minSize
maxSizeOld=maxSize
watershedOld=watershed
Thread.sleep(200)
labelPrevImp.close()
makeNearProj = gd.checkboxes.get(3).getState()
return segmentChannel, donorChannel, acceptorChannel, acceptorChannel2, thresholdMethod, maxIntensity, gaussianSigma, largeDoGSigma, topHat, topHatSigma, manualSegment, manualThreshold, makeNearProj, dilation, sizeExclude, minSize, maxSize, watershed
def overlay_curvatures(imp,
curvature_profiles,
params,
limits=None,
annotate=True):
"""Overlay curvature pixels on membrane image"""
membrane_channel = params.membrane_channel_number
overlay_base_imp = imp.clone()
curvature_stack = ImageStack(imp.getWidth(), imp.getHeight())
for profile in curvature_profiles:
curvature_stack = generate_curvature_overlays(profile, curvature_stack)
overlay_imp = ImagePlus("Curvature stack", curvature_stack)
IJ.run(overlay_imp, params.curvature_overlay_lut_string, "")
if limits is None:
flat_list_curv = [c[1] for cs in curvature_profiles for c in cs]
limits = [min(flat_list_curv),
max(flat_list_curv)]
IJ.setMinAndMax(overlay_imp, limits[0], limits[1])
raw_overlay = Duplicator().run(overlay_imp)
if annotate:
cb_fraction = 0.05
overlay_imp = add_colorbar(overlay_imp, limits, cb_fraction)
IJ.run(overlay_imp, "RGB Color", "")
w = overlay_imp.getWidth()
h = overlay_imp.getHeight()
overlaid_stack = ImageStack(w, h)
for fridx in range(1, curvature_stack.getSize() + 1):
raw_idx = overlay_base_imp.getStackIndex(membrane_channel, 1, fridx)
ip = overlay_base_imp.getStack().getProcessor(raw_idx).convertToRGB()
pix = overlay_imp.getStack().getProcessor(fridx).getPixels()
base_pix = ip.getPixels()
curv = [c for ((x, y), c) in curvature_profiles[fridx - 1]]
start_idx = next((i for i, x in enumerate(curv) if x), None)
end_idx = next(
(len(curv) - i for i, x in enumerate(reversed(curv)) if x), None)
for ((x, y), c) in curvature_profiles[fridx - 1][start_idx:end_idx]:
# ensure that no rounding issues cause pixels to fall outside image...
if x > (w - 1):
x = w - 1
if y > (h - 1):
y = h - 1
if x < 0:
x = 0
if y < 0:
y = 0
if params.filter_negative_curvatures:
if (c > 0):
base_pix[int(round(y)) * w +
int(round(x))] = pix[int(round(y)) * w +
int(round(x))]
else:
base_pix[int(round(y)) * w +
int(round(x))] = pix[int(round(y)) * w +
int(round(x))]
if annotate:
for x in range(w - int(w * cb_fraction), w):
for y in range(0, h):
base_pix[int(round(y)) * w +
int(round(x))] = pix[int(round(y)) * w +
int(round(x))]
overlaid_stack.addSlice(ip)
out_imp = ImagePlus("Overlaid curvatures", overlaid_stack)
if annotate:
out_imp = generate_limit_labels(out_imp, limits, cb_fraction, params)
FileSaver(raw_overlay).saveAsTiff(
os.path.join(params.output_path, "raw curvature.tif"))
return out_imp, raw_overlay
glVal[i] = i cts = hist[i] if(cts > maxCts): maxCts = cts totCts += cts glCum[i] = float(totCts) # print(totCts) for i in range(0, nBins): glCum[i] /= totCts frac = 1.0 n = 1 thr = 0 while(frac >= factor): thr = nBins-n frac = glCum[thr] n += 1 print(thr) IJ.setMinAndMax(imp, 0, thr)
import glob, os
__author__ = 'Nichole Wespe'
imp = IJ.getImage()
directory = IJ.getDirectory("image")
f = str(imp.getTitle())
date, rows, columns, time_tif = str.split(f) # deconstruct filename
time = time_tif.replace('tif','jpg') # need to remove .tif from time
row_IDs = list(6*rows[0] + 6*rows[1])
column_IDs = [str(i) for i in 2*(range(int(columns[0]), int(columns[0]) + 6))]
zipped = zip(row_IDs, column_IDs)
sample_names = ["".join(values) for values in zipped]
IJ.setMinAndMax(1900, 11717) # adjust brightness and contrast
IJ.run("Apply LUT")
IJ.makeRectangle(200, 50, 730, 950) # initialize selection
dial = WaitForUserDialog("Adjust selection area")
dial.show() # ask user to place selection appropriately
IJ.run("Crop")
adjDir = os.path.join(directory, "Adjusted")
if not os.path.exists(adjDir):
os.mkdir(adjDir)
adjImage = os.path.join(adjDir, "Adj " + f)
IJ.saveAs("Jpeg", adjImage)
## make ROI list
w = 130
h = 480
x_vals = 2*[10, 128, 246, 360, 478, 596]
def process(dirIn, dirOut, expName, ps, pe, ts, te):
jobid = commands.getoutput("echo $PBS_JOBID").split('.')[0]
jobid = jobid.replace("[","_").replace("]","") # because the jobids look like 2356385[1] which causes problems
print "job id: "+jobid
jobdir = os.path.join("/tmp",str(jobid)+"_fiji")
print "job dir: "+jobdir
for p in range(ps,pe+1):
pID = str(p);
for t in range(ts,te+1):
print "TIMEPOINT STARTING ***************"
tID = "t"+str(t);
print "time-point: "+tID;
if os.path.isdir(jobdir):
print "removing "+jobdir
shutil.rmtree(jobdir)
print "creating "+jobdir
os.mkdir(jobdir)
if stitching:
fileOut = "rescaled_flipped_";
for z in range(zs,ze+1):
zID = "z"+str(z);
print "z-plane: "+zID;
if bandpass:
IJ.log("bandpass....")
# load all images from same time point and same z-position
fileID = expName+pID+"_b0"+tID+zID+"m.*";
IJ.log("opening images: "+os.path.join(dirIn,fileID))
IJ.run("Image Sequence...", "open=["+dirIn+"] starting=1 increment=1 scale=100 file=[] or=["+fileID+"] sort");
#selectWindow("im-2012-0007_Position35.tif_Files");
imp = IJ.getImage();
#imp.show();
imp.setTitle("Stack");
# illumination correction
IJ.log("computing FFT...");
# run("Flip Horizontally", "stack");
impFFT = Duplicator().run(imp);
for i in range(1, impFFT.getNSlices()+1):
print "FFT of slice "+str(i)
impFFT.setSlice(i)
IJ.run(impFFT, "Bandpass Filter...", "filter_large=10000 filter_small=200 suppress=None tolerance=5 ");
#impFFT.show()
#stats = imp.getStatistics(Measurements.MEAN)
#IJ.log("stats.mean = "+str(stats.mean)); # this is only the mean of one slice...is this a problem?
print "dividing image stack by FFT stack...";
ic = ImageCalculator()
impCorr = ic.run("Divide create 32-bit stack", imp, impFFT);
#impCorr.show()
def computeMean(pixels):
return sum(pixels) / float(len(pixels))
print "multiplying each image by 128/mean for going back to 8 bit space..."
stack = impCorr.getStack()
for i in range(1, impCorr.getNSlices()+1):
ip = stack.getProcessor(i).convertToFloat()
mean = computeMean(ip.getPixels())
print "multiplying slice "+str(i)+" by "+str(float(128/mean))
ip.multiply(float(128/mean))
IJ.log("converting from 32-bit to 8-bit...")
IJ.setMinAndMax(impCorr, 0, 255);
IJ.run(impCorr,"8-bit","");
#IJ.saveAs(impCorr, "Tiff", "/Users/tischi/Documents/processed.tif");
#ff
#impCorr.show()
# save images
IJ.log("saving bandpass corrected image sequence: "+os.path.join(jobdir,fileOut))
IJ.run(impCorr, "Image Sequence... ", "format=TIFF name=["+fileOut+"] start=1 digits=4 save=["+jobdir+"]");
if check:
IJ.run(impCorr, "Image Sequence... ", "format=TIFF name=["+fileOut+"] start=1 digits=4 save=["+dirOut+"]");
#impCorr.close(); imp.close(); impFFT.hide();
# stitching
IJ.log("STITCHING START **********")
layoutFile = copyStitchingLayoutFile(dirIn,expName,jobdir,ps)
###layoutFile = makeStitchingLayoutFile(jobdir)
createPreview = 0
computeOverlap = 0
fusion_method="Linear Blending"
handleRGB = "Red, Green and Blue"
showImage = 0
#fusion=1 regression=0.30 max/avg=2.50 absolute=3.50"
st = Stitch_Image_Collection()
st.alpha = 1
IJ.log("layout file: "+str(layoutFile))
impStitched = st.work(layoutFile, createPreview, computeOverlap, fusion_method, handleRGB, showImage)
stitchedFile = os.path.join(jobdir,tID+zID+"_stitched.tif");
#impStitched.show()
IJ.saveAs(impStitched,"Tiff", stitchedFile);
if check:
print os.path.join(dirOut,tID+zID+"_stitched.tif")
stitchedFile = os.path.join(dirOut,tID+zID+"_stitched.tif");
IJ.saveAs(impStitched,"Tiff", stitchedFile);
IJ.log("STITCHING END **********")
if combine_z:
IJ.log("combine z.....")
#########
IJ.log("load stitched images into a stack...")
for z in range(zs,ze+1):
zID = "z"+str(z);
stitchedFile = os.path.join(jobdir,tID+zID+"_stitched.tif");
IJ.log("opening "+stitchedFile)
imp = IJ.openImage(stitchedFile)
if z==zs:
stack = ImageStack(imp.width,imp.height)
stack.addSlice(imp.getProcessor())
imp = ImagePlus("stack", stack)
#imp.show()
########
########
IJ.log("cropping...")
imp.setRoi(xs, ys, xe, ye);
IJ.run(imp, "Crop", "");
#imp.show()
########
# the following normalisation should not be necessary, because they are already all 128/mean normalised
#IJ.log("-- normalise intensity of all slices...")
#stats = imp.getStatistics(Measurements.MEAN)
#IJ.log("stats.mean = "+str(stats.mean)); # this is only the mean of one slice...is this a problem?
#stack = imp.getStack()
#for i in range(1, impFFT.getNSlices()+1):
# ip = stack.getProcessor(i).convertToFloat()
# ip.multiply(128/stats.mean)
# #stack.setSlice(stack.getSliceLabel(i), ip)
#run("Set Slice...", "slice="+1);
#run("Set Measurements...", " mean redirect=None decimal=9");
#run("Select None");
#setBatchMode(true);
#setMinAndMax(0, 255); run("32-bit");
#for(l=0; l<nSlices+1; l++) {
## run("Select All");
# run("Clear Results");
# run("Measure");
# picsum=getResult("Mean",0);
# //if(l==0){picsum1=picsum;}
# //int_ratio=picsum1/picsum;
# int_ratio=128/picsum;
# run("Select None");
# IJ.log("ratio ="+int_ratio);
# run("Multiply...", "slice value="+int_ratio);
# run("Next Slice [>]");
#}
#setBatchMode(false);
# stop here and try by hand
#...
#dfgfd
#//stack-reg
#//IJ.log("register xy...")
#//run("StackReg", "transformation=Translation");
#// project into 1 plane ....
#// run("Extended Depth of Field (Easy mode)...");
#//run("Z Project...", "start=["+1+"] stop=["+ze+"] projection=[Standard Deviation]");
doEDF = True
if doEDF:
IJ.log("EDF: start...")
parameters = Parameters()
parameters.setQualitySettings(1)
parameters.setTopologySettings(0)
parameters.show3dView = False
parameters.showTopology = False
edfh = ExtendedDepthOfFieldHeadless(imp, parameters)
imp = edfh.processHeadless()
IJ.log("EDF: done.")
#imp.show()
IJ.log("EDF: converting from 32-bit to 8-bit...")
IJ.setMinAndMax(imp, 0, 255);
IJ.run(imp,"8-bit","");
edfFile = os.path.join(dirOut,expName+pID+"-"+tID+"_EDOF_noTimeNorm.tif");
IJ.log("EDF save: "+edfFile)
IJ.saveAs(imp,"Tiff", edfFile);
IJ.log("EDF save: done.")
#close(); // projection
#close(); // stack
print "TIMEPOINT FINISHED ***************"
if os.path.isdir(jobdir):
print "removing "+jobdir
shutil.rmtree(jobdir)
imp.setTitle("current")
# IJ.run executes commands from the macro recorder
IJ.run(
"Scale...",
"x=.5 y=.5 z=1.0 width=128 height=128 depth=930 interpolation=Bilinear average process create"
)
imp.close()
IJ.selectWindow("current-1")
IJ.run("Z Project...", "projection=[Average Intensity]")
# Note the new imports above, WindowManager and ImageCalculator
# ic.run executes commands within image calculator; wm.getImage selects the image using WindowsManager
imp = ic.run("Subtract create 32-bit stack", wm.getImage("current-1"),
wm.getImage("AVG_current-1"))
imp.show()
imp = ic.run("Divide create 32-bit stack", wm.getImage("Result of current-1"),
wm.getImage("AVG_current-1"))
imp.show()
IJ.selectWindow("Result of Result of current-1")
# adjust the Brightness/Contrast
IJ.setMinAndMax(-0.0200, 0.2000)
# this is the Lookup Tables step
IJ.run("Aselfmade3")
# IJ.saveAs has 3 arguments: (image, file type, file path)
IJ.saveAs(IJ.getImage(), "Tiff", out_folder + the_file + '-processed')
IJ.run("Close All", "")
source_path = "Correspondence_ZBrain/ZBtoEM/ZBdata_warped/"
reformat_path = "Correspondence_ZBrain/ZBtoEM/ZBdata_imseq/"
for filename in os.listdir(source_path):
if filename.endswith(".tif"):
print "Processing file:", filename
## Open image from file
imp = IJ.openImage(os.path.join(source_path, filename))
if imp is None:
print "Could not open image from file:", filename
continue
## Set appropriate resolution (matching BigWarp settings here)
IJ.run(imp, "Properties...", "channels=1 slices=811 frames=1 unit=um pixel_width=0.6 pixel_height=0.6 voxel_depth=1.2")
## Make sure full 16-bit or 8-bit range is used
IJ.setMinAndMax(imp, 0, 65535) # ZBrain/ZBB data stacks
imp.setDefault16bitRange(16)
#IJ.setMinAndMax(imp, 0, 255); # ZBrain masks/labels
#imp.setDefault16bitRange(8);
## Convert to 8-bit (optional, if desired)
IJ.run(imp, "8-bit", "")
## If converted, make sure full 8-bit range is used before saving
IJ.setMinAndMax(imp, 0, 255);
imp.setDefault16bitRange(8);
fs = FileSaver(imp)
if not path.exists(reformat_path):
os.makedirs(reformat_path)
if path.exists(reformat_path) and path.isdir(reformat_path):
dirpath = path.join(reformat_path, string.replace(filename, '_Warped.tif', ''))
if not path.exists(dirpath):
