def correct_drift(img1, img2, display_cc=False):
''' Returns two ImagePlus objects that are drift corrected to each other.
The first image is not changed.
The second image is a new image that is shifted using ImageJ's Translate.
:param img1: The reference image.
:param img2: The image to be shifted.
:param display_cc: Activate displaying the CrossCorrelation image (default False).
'''
img1_cc, img2_cc = cc.scale_to_power_of_two([img1, img2])
result = cc.perform_correlation(img1_cc, img2_cc)
x_off, y_off = cc.get_shift(result)
# style after maximum detection
if not display_cc:
result.hide()
else:
result.copyScale(img1)
cc.style_cc(result)
if x_off == 0 and y_off == 0:
print('No drift has been detected.')
return img1, img2
title = img2.getTitle()
img2_dk = Duplicator().run(img2)
img2_dk.setTitle('DK-' + title)
IJ.run(img2_dk, 'Translate...', 'x=%d y=%d interpolation=None' % (-x_off, -y_off))
img2_dk.copyScale(img2)
img2_dk.show()
return img1, img2_dk
def threshold(imPlus, edgeThreshold=2500):
mask = Duplicator().run(imPlus)
mask_stk = mask.getStack()
# First, we threshold based on edges
IJ.setThreshold(mask, edgeThreshold, 100000, "No Update")
for i in range(mask.getImageStackSize()):
mask_stk.getProcessor(i + 1).findEdges()
IJ.run(mask, "Make Binary", "method=Default background=Default black")
# Now, we need to clean up the binary images morphologically
IJ.run(mask, "Dilate", "stack")
IJ.run(mask, "Fill Holes", "stack")
IJ.run(mask, "Erode", "stack")
IJ.run(mask, "Erode", "stack")
# Finally, remove the small particles
stk = ImageStack(mask.getWidth(), mask.getHeight())
p = PA(PA.SHOW_MASKS, 0, None, 200, 100000)
p.setHideOutputImage(True)
for i in range(mask_stk.getSize()):
mask.setSliceWithoutUpdate(i + 1)
p.analyze(mask)
mmap = p.getOutputImage()
stk.addSlice(mmap.getProcessor())
mask.setStack(stk)
mask.setSliceWithoutUpdate(1)
mask.setTitle(mask_title(imPlus.getTitle()))
mask.show()
return mask
def blurImage(ip):
#duplicate
imp_dup = Duplicator().run(ip)
#blur it
imp_dup.show()
IJ.run(imp_dup, "Gaussian Blur...", "sigma=1")
imp_GB = IJ.getImage()
return (imp_GB)
def z_crop(imp):
"""trim a z stack based on interactively-defined start and end points"""
WaitForUserDialog("Choose first z plane and click OK...").show()
start_z = imp.getZ()
WaitForUserDialog("Now choose last z plane and click OK...").show()
end_z = imp.getZ()
frames = imp.getNFrames()
channels = imp.getNChannels()
#current_channel = imp.getC();
dupimp = Duplicator().run(imp, 1, channels, start_z, end_z, 1, frames)
imp.changes = False
imp.close()
dupimp.show()
#autoset_zoom(dupimp);
return dupimp, (start_z, end_z)
def z_crop(imp):
"""trim a z stack based on interactively-defined start and end points"""
IJ.setTool("zoom")
IJ.run("Brightness/Contrast...")
imp.setZ(1)
WaitForUserDialog("Choose first z plane and click OK...").show()
start_z = imp.getZ()
WaitForUserDialog("Now choose last z plane and click OK...").show()
end_z = imp.getZ()
frames = imp.getNFrames()
channels = imp.getNChannels()
imp.killRoi()
dupimp = Duplicator().run(imp, 1, channels, start_z, end_z, 1, frames)
imp.hide()
dupimp.show()
return dupimp, (start_z, end_z)
def cut(event):
roi = imp.getRoi()
if roi != None:
newRoi = roi.clone()
Dup = Duplicator().run(imp, 1, imp.getNChannels(), 1, imp.getNSlices(), 1, imp.getNFrames())
newRoi.setLocation(0,0)
Dup.setRoi(newRoi)
Dup.setTitle(Men.getTextField() + str(Men.getCounter()))
Dup.show()
#Men.setCounter()
Men.addOlay(roi)
imp.setOverlay(Men.getOverlay()) #setOverlay(Roi roi, java.awt.Color strokeColor, int strokeWidth, java.awt.Color fillColor)
imp.getOverlay().drawLabels(True) # drawNumbers
imp.deleteRoi()
#make cell instance and add to position instance
p.addCell(cell(p.getMainPath(), p, Men.getCounter(), Dup)) # cell(mainPath, position, ID, imp):
Dup.close()
Men.increaseCounter()
distMin = dist
return centPart, (x, y)
def isSpecialParticle(image, roim, iParticle, boundariesNuc, boundariesMito, pixelSize, z):
try:
image.setSlice(int(round(z)))
image2 = ImagePlus("slice",image.getProcessor())
roim.select(image2, iParticle)
image3 = Duplicator().run(image2);
image3.show()
sp = segment_Particles()
image3 = sp.preprocessImage(image3, "MAX", "C1", 0, 0)
image3 = sp.segmentParticles(image3, "Triangle", boundariesNuc[0], boundariesNuc[1], 0, 0)
if roim.getCount() > 0:
roim.select(image3, 0)
stats = image3.getStatistics(Measurements.AREA + Measurements.SHAPE_DESCRIPTORS)
IJ.log("Area (um2) " + str(stats.area*pixelSize*pixelSize) + "; Aspect Ratio " + str(stats.major/stats.minor))
if stats.area > boundariesMito[0] and stats.area < boundariesMito[1] and stats.major/stats.minor > boundariesMito[2]:
return 1
else:
return 0
except BaseException, err:
def qc_background_regions(intensity_imp, bg_rois):
"""allow the user to view and correct automatically-determined background regions"""
imp = Duplicator().run(intensity_imp);
imp.setTitle("Background region QC");
imp.show();
imp.setPosition(1);
autoset_zoom(imp);
imp.setRoi(bg_rois[0]);
IJ.setTool("freehand");
notOK = True;
while notOK:
listener = UpdateRoiImageListener(bg_rois, is_area=True);
imp.addImageListener(listener);
dialog = NonBlockingGenericDialog("Background region quality control");
dialog.enableYesNoCancel("Continue", "Use this region for all t");
dialog.setCancelLabel("Cancel analysis");
dialog.addMessage("Please redraw background regions as necessary...")
dialog.showDialog();
if dialog.wasCanceled():
raise KeyboardInterrupt("Run canceled");
elif not(dialog.wasOKed()):
this_roi = imp.getRoi();
bg_rois = [this_roi for _ in listener.getRoiList()];
imp.removeImageListener(listener);
else:
last_roi = imp.getRoi();
qcd_bg_rois = listener.getRoiList();
if imp.getNFrames() > imp.getNSlices():
qcd_bg_rois[imp.getT() - 1] = last_roi;
else:
qcd_bg_rois[imp.getZ() - 1] = last_roi;
notOK = False;
imp.removeImageListener(listener);
imp.changes = False;
imp.close();
return qcd_bg_rois;
def time_crop(imp, params):
"""trim a time series based on interactively-defined start and end points"""
start_frame = None;
end_frame = None;
if params.perform_time_crop:
if params.time_crop_start_end is not None:
start_frame = params.time_crop_start_end[0];
end_frame = params.time_crop_start_end[1];
else:
MyWaitForUser("First T...", "Choose first time frame and click OK...");
start_frame = imp.getT();
MyWaitForUser("Last T...", "Now choose last time frame and click OK...");
end_frame = imp.getT();
slices = imp.getNSlices();
channels = imp.getNChannels();
if start_frame is not None:
dupimp = Duplicator().run(imp, 1, channels, 1, slices, start_frame, end_frame);
imp.changes = False;
imp.close();
dupimp.show()
autoset_zoom(dupimp);
return dupimp, (start_frame, end_frame);
else:
return imp, (start_frame, end_frame);
class gui(JFrame):
def __init__(self): # constructor
#origing of coordinates
self.coordx = 10
self.coordy = 10
#inintialize values
self.Canvas = None
#create panel (what is inside the GUI)
self.panel = self.getContentPane()
self.panel.setLayout(GridLayout(9, 2))
self.setTitle('Subdividing ROIs')
#define buttons here:
self.Dapi_files = DefaultListModel()
mylist = JList(self.Dapi_files, valueChanged=self.open_dapi_image)
#mylist.setSelectionMode(ListSelectionModel.SINGLE_SELECTION);
mylist.setLayoutOrientation(JList.VERTICAL)
mylist.setVisibleRowCount(-1)
listScroller1 = JScrollPane(mylist)
listScroller1.setPreferredSize(Dimension(300, 80))
self.output_files = DefaultListModel()
mylist2 = JList(self.output_files)
#mylist.setSelectionMode(ListSelectionModel.SINGLE_SELECTION);
mylist2.setLayoutOrientation(JList.VERTICAL)
mylist2.setVisibleRowCount(-1)
listScroller2 = JScrollPane(mylist2)
listScroller2.setPreferredSize(Dimension(300, 80))
quitButton = JButton("Quit", actionPerformed=self.quit)
selectInputFolderButton = JButton("Select Input",
actionPerformed=self.select_input)
cubifyROIButton = JButton("Cubify ROI",
actionPerformed=self.cubify_ROI)
saveButton = JButton("Save ROIs", actionPerformed=self.save)
summsaveButton = JButton("Save Summary",
actionPerformed=self.save_summary)
self.textfield1 = JTextField('500')
#add buttons here
self.panel.add(listScroller1)
self.panel.add(listScroller2)
self.panel.add(selectInputFolderButton)
self.panel.add(Label("Adjust the size of the ROIs"))
self.panel.add(self.textfield1)
self.panel.add(cubifyROIButton)
self.panel.add(saveButton)
self.panel.add(summsaveButton)
self.panel.add(quitButton)
#self.panel.add(saveButton)
#self.panel.add(Zslider)
#other stuff to improve the look
self.pack() # packs the frame
self.setVisible(True) # shows the JFrame
self.setLocation(self.coordx, self.coordy)
#define functions for the buttons:
def quit(self, event): #quit the gui
self.dispose()
IJ.run("Close All")
def select_input(self, event):
self.input_path = IJ.getDirectory(
"Select Directory containing your data")
# get the files in that directory
self.input_files = listdir(self.input_path)
self.input_dapi = [s for s in self.input_files if "DAPI.tif" in s]
self.core_names = get_core_names(self.input_dapi)
# create output directory
self.output_path = path.join(
path.dirname(path.dirname(self.input_path)), "Cubified_ROIs")
if path.isdir(self.output_path):
print "Output path already created"
else:
mkdir(self.output_path)
print "Output path created"
# get the processed data
self.processed_files = listdir(self.output_path)
self.out_core_names = get_core_names(
get_core_names(self.processed_files)
) # needs the channel and roi to be removed
# populate the lists
for f in set(self.core_names):
if f not in set(self.out_core_names): # skip if processed
self.Dapi_files.addElement(f)
for f in set(self.out_core_names
): # a set so that only unique ones are shown
self.output_files.addElement(f)
def open_dapi_image(self, e):
sender = e.getSource()
IJ.run("Close All")
if not e.getValueIsAdjusting():
self.name = sender.getSelectedValue()
print self.name
dapi_filename = path.join(self.input_path, self.name + '_DAPI.tif')
#print dapi_filename
if not path.exists(dapi_filename):
print "I don't find the DAPI file, which is weird as I just found it before"
else:
#open stack and make the reslice
self.dapi_image = ImagePlus(dapi_filename) #read the image
# duplicate image to play with that one, and do the real processing in the background
self.imp_main = Duplicator().run(self.dapi_image)
self.imp_main.setTitle(self.name)
self.imp_main.show() #show image on the left
#self.original_image.getWindow().setLocation(self.coordx-self.original_image.getWindow().getWidth()-10,self.coordy-10) #reposition image
def cubify_ROI(self, e):
self.L = int(self.textfield1.text)
# get info
tit = self.imp_main.getTitle()
self.roi = self.imp_main.getRoi()
# get corners
corners = get_corners(self.roi, self.L)
self.corners_cleaned = clean_corners(corners, self.roi, self.L)
print 'found corners'
# get the overlay
self.ov = overlay_corners(self.corners_cleaned, self.L)
self.ov = overlay_roi(self.roi, self.ov)
# write roi name
self.ov = write_roi_numbers(self.ov, self.corners_cleaned, self.L)
# overlay
self.imp_main.setOverlay(self.ov)
self.imp_main.updateAndDraw()
def save(self, e):
# open the other channels
d1_filename = path.join(self.input_path, self.name + '_D1.tif')
d2_filename = path.join(self.input_path, self.name + '_D2.tif')
self.d1_image = ImagePlus(d1_filename) #read the image
print "d1 image opened"
self.d2_image = ImagePlus(d2_filename) #read the image
print "d2 image opened"
for image in [self.dapi_image, self.d1_image, self.d2_image]:
print "saving rois for image " + image.getTitle()
save_rois(image, self.corners_cleaned, self.L, self.output_path)
print "ROIs saved"
def save_summary(self, e):
IJ.selectWindow(self.name)
IJ.run("Flatten")
imp = IJ.getImage()
# downsample
# scale ENLARGING or SHRINKING the canvas dimensions
scale_factor = .2
new_width = str(int(imp.getWidth() * scale_factor))
new_height = str(int(imp.getHeight() * scale_factor))
IJ.selectWindow(self.name + "-1")
str_to_scale = "x=" + str(scale_factor) + " y=" + str(
scale_factor
) + " width=" + new_width + " height=" + new_height + " interpolation=Bilinear average create"
IJ.run("Scale...", str_to_scale)
# save
imp2 = IJ.getImage()
#IJ.saveAs("Tiff", path.join(OUTDIR, self.name + "_summaryOfROIs"))
IJ.saveAsTiff(
imp2, path.join(self.output_path,
self.name + "_summaryOfROIs.tif"))
print "summary image saved"
######### open image using dialogue box
#imp = IJ.getImage()
original = IJ.openImage(getFile())
original.show()
########## Use thresholding and selection to define UFOV ###################################################################################
#IJ.run("ROI Manager...", "") # not sure if I need this
IJ.setRawThreshold(original, 1, 255,'') # background pixels have value 0. See IMAGE>ADJUST>THRESHOLD
IJ.run(original, "Create Selection", "") # add bounding box. See EDIT>SELECTION
IJ.run(original,"To Bounding Box", "") # this box defines the UFOV. See EDIT>SELECTION
IJ.resetThreshold(original) # get back original now UFOV is definedthresholding
UFOV = Duplicator().run(original) # duplicate the original image, only the CFOV
UFOV.setTitle("UFOV")
UFOV.show()
CFOV_fraction = 0.75 # choose the fraction of the UFOV that defines the CFOV
IJ.run(original,"Scale... ", "x="+str(CFOV_fraction)+" y="+str(CFOV_fraction)+" centered") # rescale bounding box to get CFOV
CFOV = Duplicator().run(original) # duplicate the original image, only the CFOV
CFOV.setTitle("CFOV")
CFOV.show()
######### Nema process including Re-bin image to larger pixels ################################################################################
desired_pixel_width = getPixel() # 6.4 mm default, remember tolerance is +/-30%
current_pixel_width = CFOV.getCalibration().pixelWidth #get pixel width, 1.16 mm
shrink_factor = int(desired_pixel_width/current_pixel_width) # must be an integer
IJ.run(CFOV, "Bin...", "x="+str(shrink_factor)+" y="+str(shrink_factor)+" bin=Sum") # run the bin plugin
def runOneFile(fullFilePath):
global gFileType
global gNumChannels
global gAlignBatchVersion
if not os.path.isfile(fullFilePath):
bPrintLog('\nERROR: runOneFile() did not find file: ' + fullFilePath + '\n',0)
return 0
bPrintLog(time.strftime("%H:%M:%S") + ' starting runOneFile(): ' + fullFilePath, 1)
enclosingPath = os.path.dirname(fullFilePath)
head, tail = os.path.split(enclosingPath)
enclosingPath += '/'
#make output folders
destFolder = enclosingPath + tail + '_channels/'
if not os.path.isdir(destFolder):
os.makedirs(destFolder)
destMaxFolder = destFolder + 'max/'
if not os.path.isdir(destMaxFolder):
os.makedirs(destMaxFolder)
if gDoAlign:
destAlignmentFolder = destFolder + 'alignment/'
if not os.path.isdir(destAlignmentFolder):
os.makedirs(destAlignmentFolder)
if gSave8bit:
eightBitFolder = destFolder + 'channels8/'
if not os.path.isdir(eightBitFolder):
os.makedirs(eightBitFolder)
eightBitMaxFolder = eightBitFolder + 'max/'
if not os.path.isdir(eightBitMaxFolder):
os.makedirs(eightBitMaxFolder)
if gFileType=='tif':
# open .tif image
imp = Opener().openImage(fullFilePath)
else:
# open .lsm
cmdStr = 'open=%s autoscale color_mode=Default view=Hyperstack stack_order=XYCZT' % (fullFilePath,)
IJ.run('Bio-Formats Importer', cmdStr)
lsmpath, lsmfilename = os.path.split(fullFilePath)
lsWindow = lsmfilename
imp = WindowManager.getImage(lsWindow)
# get parameters of image
(width, height, nChannels, nSlices, nFrames) = imp.getDimensions()
bitDepth = imp.getBitDepth()
infoStr = imp.getProperty("Info") #get all .tif tags
if not infoStr:
infoStr = ''
infoStr += 'bAlignBatch_Version=' + str(gAlignBatchVersion) + '\n'
infoStr += 'bAlignBatch_Time=' + time.strftime("%Y%m%d") + '_' + time.strftime("%H%M%S") + '\n'
msgStr = 'w:' + str(width) + ' h:' + str(height) + ' slices:' + str(nSlices) \
+ ' channels:' + str(nChannels) + ' frames:' + str(nFrames) + ' bitDepth:' + str(bitDepth)
bPrintLog(msgStr, 1)
path, filename = os.path.split(fullFilePath)
shortName, fileExtension = os.path.splitext(filename)
#
# look for num channels in ScanImage infoStr
if gGetNumChanFromScanImage:
for line in infoStr.split('\n'):
#scanimage.SI4.channelsSave = [1;2]
scanimage4 = find(line, 'scanimage.SI4.channelsSave =') == 0
#state.acq.numberOfChannelsSave=2
scanimage3 = find(line, 'state.acq.numberOfChannelsSave=') == 0
if scanimage3:
#print 'line:', line
equalIdx = find(line, '=')
line2 = line[equalIdx+1:]
if gGetNumChanFromScanImage:
gNumChannels = int(line2)
bPrintLog('over-riding gNumChannels with: ' + str(gNumChannels), 2)
if scanimage4:
#print ' we have a scanimage 4 file ... now i need to exptract the number of channel'
#print 'line:', line
equalIdx = find(line, '=')
line2 = line[equalIdx+1:]
for delim in ';[]':
line2 = line2.replace(delim, ' ')
if gGetNumChanFromScanImage:
gNumChannels = len(line2.split())
bPrintLog('over-riding gNumChannels with: ' + str(gNumChannels), 2)
# show
imp.show()
# split channels if necc. and grab the original window names
if gNumChannels == 1:
origImpWinStr = imp.getTitle() #use this when only one channel
origImpWin = WindowManager.getWindow(origImpWinStr) #returns java.awt.Window
if gNumChannels == 2:
winTitle = imp.getTitle()
bPrintLog('Deinterleaving 2 channels...', 1)
IJ.run('Deinterleave', 'how=2 keep') #makes ' #1' and ' #2', with ' #2' frontmost
origCh1WinStr = winTitle + ' #1'
origCh2WinStr = winTitle + ' #2'
origCh1Imp = WindowManager.getImage(origCh1WinStr)
origCh2Imp = WindowManager.getImage(origCh2WinStr)
origCh1File = destFolder + shortName + '_ch1.tif'
origCh2File = destFolder + shortName + '_ch2.tif'
# work on a copy, mostly for alignment with cropping
copy = Duplicator().run(imp)
#copy.copyAttributes(imp) #don't copy attributes, it copies the name (which we do not want)
copy.show()
#
# crop (on copy)
if gDoCrop:
bPrintLog('making cropping rectangle (left,top,width,height) ',1)
bPrintLog(str(gCropLeft) + ' ' + str(gCropTop) + ' ' +str(gCropWidth) + ' ' +str(gCropHeight), 2)
roi = Roi(gCropLeft, gCropTop, gCropWidth, gCropHeight) #left,top,width,height
copy.setRoi(roi)
time.sleep(0.5) # otherwise, crop SOMETIMES failes. WHAT THE F**K FIJI DEVELOPERS, REALLY, WHAT THE F**K
#bPrintLog('cropping', 1)
IJ.run('Crop')
infoStr += 'bCropping=' + str(gCropLeft) + ',' + str(gCropTop) + ',' + str(gCropWidth) + ',' + str(gCropHeight) + '\n'
#
# remove calibration ( on original)
if gRemoveCalibration:
cal = imp.getCalibration()
calCoeff = cal.getCoefficients()
if calCoeff:
msgStr = 'Calibration is y=a+bx' + ' a=' + str(calCoeff[0]) + ' b=' + str(calCoeff[1])
bPrintLog(msgStr, 1)
#remove calibration
bPrintLog('\tRemoving Calibration', 2)
imp.setCalibration(None)
#without these, 8-bit conversion goes to all 0 !!! what the f**k !!!
#bPrintLog('calling imp.resetStack() and imp.resetDisplayRange()', 2)
imp.resetStack()
imp.resetDisplayRange()
#get and print out min/max
origMin = StackStatistics(imp).min
origMax = StackStatistics(imp).max
msgStr = '\torig min=' + str(origMin) + ' max=' + str(origMax)
bPrintLog(msgStr, 2)
# 20150723, 'shift everybody over by linear calibration intercept calCoeff[0] - (magic number)
if 1:
# [1] was this
#msgStr = 'Subtracting original min '+str(origMin) + ' from stack.'
#bPrintLog(msgStr, 2)
#subArgVal = 'value=%s stack' % (origMin,)
#IJ.run('Subtract...', subArgVal)
# [2] now this
#msgStr = 'Adding calCoeff[0] '+str(calCoeff[0]) + ' from stack.'
#bPrintLog(msgStr, 2)
#addArgVal = 'value=%s stack' % (int(calCoeff[0]),)
#IJ.run('Add...', addArgVal)
# [3] subtract a magic number 2^15-2^7 = 32768 - 128
magicNumber = gLinearShift #2^15 - 128
msgStr = 'Subtracting a magic number (linear shift) '+str(magicNumber) + ' from stack.'
bPrintLog(msgStr, 2)
infoStr += 'bLinearShift=' + str(gLinearShift) + '\n'
subArgVal = 'value=%s stack' % (gLinearShift,)
IJ.run(imp, 'Subtract...', subArgVal)
# 20150701, set any pixel <0 to 0
if 0:
ip = imp.getProcessor() # returns a reference
pixels = ip.getPixels() # returns a reference
msgStr = '\tSet all pixels <0 to 0. This was added 20150701 ...'
bPrintLog(msgStr, 2)
pixels = map(lambda x: 0 if x<0 else x, pixels)
bPrintLog('\t\t... done', 2)
#get and print out min/max
newMin = StackStatistics(imp).min
newMax = StackStatistics(imp).max
msgStr = '\tnew min=' + str(newMin) + ' max=' + str(newMax)
bPrintLog(msgStr, 2)
#append calibration to info string
infoStr += 'bCalibCoeff_a = ' + str(calCoeff[0]) + '\n'
infoStr += 'bCalibCoeff_b = ' + str(calCoeff[1]) + '\n'
infoStr += 'bNewMin = ' + str(newMin) + '\n'
infoStr += 'bNewMax = ' + str(newMax) + '\n'
#
# set up
if gNumChannels == 1:
impWinStr = copy.getTitle() #use this when only one channel
impWin = WindowManager.getWindow(impWinStr) #returns java.awt.Window
if gNumChannels == 2:
winTitle = copy.getTitle()
bPrintLog('Deinterleaving 2 channels...', 1)
IJ.run('Deinterleave', 'how=2 keep') #makes ' #1' and ' #2', with ' #2' frontmost
ch1WinStr = winTitle + ' #1'
ch2WinStr = winTitle + ' #2'
ch1Imp = WindowManager.getImage(ch1WinStr)
ch2Imp = WindowManager.getImage(ch2WinStr)
ch1File = destFolder + shortName + '_ch1.tif'
ch2File = destFolder + shortName + '_ch2.tif'
#
# alignment
if gDoAlign and gNumChannels == 1 and copy.getNSlices()>1:
infoStr += 'AlignOnChannel=1' + '\n'
#snap to middle slice
if gAlignOnMiddleSlice:
middleSlice = int(math.floor(copy.getNSlices() / 2)) #int() is necc., python is f*****g picky
else:
middleSlice = gAlignOnThisSlice
copy.setSlice(middleSlice)
transformationFile = destAlignmentFolder + shortName + '.txt'
bPrintLog('MultiStackReg aligning:' + impWinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=Align file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body] save' %(impWin,transformationFile)
IJ.run('MultiStackReg', stackRegParams)
infoStr += 'AlignOnSlice=' + str(middleSlice) + '\n'
#20150723, we just aligned on a cropped copy, apply alignment to original imp
origImpTitle = imp.getTitle()
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' %(origImpTitle,transformationFile)
IJ.run('MultiStackReg', stackRegParams)
if gDoAlign and gNumChannels == 2 and ch1Imp.getNSlices()>1 and ch2Imp.getNSlices()>1:
#apply to gAlignThisChannel
alignThisWindow = ''
applyAlignmentToThisWindow = ''
if gAlignThisChannel == 1:
infoStr += 'AlignOnChannel=1' + '\n'
transformationFile = destAlignmentFolder + shortName + '_ch1.txt'
alignThisWindow = ch1WinStr
applyAlignmentToThisWindow = ch2WinStr
else:
infoStr += 'AlignOnChannel=2' + '\n'
transformationFile = destAlignmentFolder + shortName + '_ch2.txt'
alignThisWindow = ch2WinStr
applyAlignmentToThisWindow = ch1WinStr
alignThisImp = WindowManager.getImage(alignThisWindow)
#snap to middle slice
if gAlignOnMiddleSlice:
middleSlice = int(math.floor(alignThisImp.getNSlices() / 2)) #int() is necc., python is f*****g picky
else:
middleSlice = gAlignOnThisSlice
alignThisImp.setSlice(middleSlice)
infoStr += 'bAlignOnSlice=' + str(middleSlice) + '\n'
bPrintLog('MultiStackReg aligning:' + alignThisWindow, 1)
stackRegParams = 'stack_1=[%s] action_1=Align file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body] save' %(alignThisWindow,transformationFile)
IJ.run('MultiStackReg', stackRegParams)
# 20150723, we just aligned on a copy, apply alignment to both channels of original
# ch1
bPrintLog('MultiStackReg applying alignment to:' + origCh1WinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' %(origCh1WinStr,transformationFile)
IJ.run('MultiStackReg', stackRegParams)
# ch2
bPrintLog('MultiStackReg applying alignment to:' + origCh2WinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' %(origCh2WinStr,transformationFile)
IJ.run('MultiStackReg', stackRegParams)
#apply alignment to other window
#bPrintLog('MultiStackReg applying alignment to:' + applyAlignmentToThisWindow, 1)
#applyAlignThisImp = WindowManager.getImage(applyAlignmentToThisWindow)
#stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' %(applyAlignmentToThisWindow,transformationFile)
#IJ.run('MultiStackReg', stackRegParams)
elif gDoAlign:
bPrintLog('Skipping alignment, there may be only one slice?',3)
#
# save
if gNumChannels == 1:
imp.setProperty("Info", infoStr);
impFile = destFolder + shortName + '.tif'
#bPrintLog('Saving:' + impFile, 1)
bSaveStack(imp, impFile)
#max project
bSaveZProject(imp, destMaxFolder, shortName)
if gNumChannels == 2:
#ch1
origCh1Imp.setProperty("Info", infoStr);
#bPrintLog('Saving:' + ch1File, 1)
bSaveStack(origCh1Imp, ch1File)
#max project
bSaveZProject(origCh1Imp, destMaxFolder, shortName+'_ch1')
#ch2
origCh2Imp.setProperty("Info", infoStr);
#bPrintLog('Saving:' + ch2File, 1)
bSaveStack(origCh2Imp, ch2File)
#max project
bSaveZProject(origCh2Imp, destMaxFolder, shortName+'_ch2')
#
# post convert to 8-bit and save
if gSave8bit:
if bitDepth == 16:
if gNumChannels == 1:
bPrintLog('Converting to 8-bit:' + impWinStr, 1)
IJ.selectWindow(impWinStr)
#IJ.run('resetMinAndMax()')
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(imp, impFile)
#max project
bSaveZProject(imp, eightBitMaxFolder, shortName)
if gNumChannels == 2:
#
bPrintLog('Converting to 8-bit:' + origCh1WinStr, 1)
IJ.selectWindow(origCh1WinStr)
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch1.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(origCh1Imp, impFile)
#max project
bSaveZProject(origCh1Imp, eightBitMaxFolder, shortName+'_ch1')
#
bPrintLog('Converting to 8-bit:' + origCh2WinStr, 1)
IJ.selectWindow(origCh2WinStr)
#IJ.run('resetMinAndMax()')
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch2.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(origCh2Imp, impFile)
#max project
bSaveZProject(origCh2Imp, eightBitMaxFolder, shortName+'_ch2')
#
# close original window
imp.changes = 0
imp.close()
#copy
copy.changes = 0
copy.close()
#
# close ch1/ch2
if gNumChannels == 2:
#original
origCh1Imp.changes = 0
origCh1Imp.close()
origCh2Imp.changes = 0
origCh2Imp.close()
#copy
ch1Imp.changes = 0
ch1Imp.close()
ch2Imp.changes = 0
ch2Imp.close()
bPrintLog(time.strftime("%H:%M:%S") + ' finished runOneFile(): ' + fullFilePath, 1)
def do_angular_projection(imp,
max_r_pix=60,
min_r_pix=10,
generate_roi_stack=True):
"""perform ray-based projection of vessel wall, c.f. ICY TubeSkinner (Lancino 2018)"""
Prefs.blackBackground = True
print("do angular projection input imp = " + str(imp))
split_chs = ChannelSplitter().split(imp)
mch_imp = split_chs[0]
IJ.setAutoThreshold(mch_imp, "IsoData dark stack")
egfp_imp = split_chs[1]
proj_imp = Duplicator().run(egfp_imp)
cl_imp = split_chs[2]
if generate_roi_stack:
egfp_imp_disp = Duplicator().run(egfp_imp)
roi_stack = IJ.createImage("rois", egfp_imp.getWidth(),
egfp_imp.getHeight(), egfp_imp.getNSlices(),
16)
centres = []
projected_im_pix = []
ring_rois = []
for zidx in range(cl_imp.getNSlices()):
if ((zidx + 1) % 100) == 0:
print("Progress = " +
str(round(100 * (float(zidx + 1) / cl_imp.getNSlices()))))
projected_im_row = []
proj_imp.setZ(zidx + 1)
mch_imp.setZ(zidx + 1)
bp = mch_imp.createThresholdMask()
bp.dilate()
bp.erode()
bp.erode()
bp.erode()
mask_imp = ImagePlus("mask", bp)
IJ.run(mask_imp, "Create Selection", "")
roi = mask_imp.getRoi()
proj_imp.setRoi(roi)
IJ.run(proj_imp, "Set...", "value=0 slice")
IJ.run(proj_imp, "Make Inverse", "")
roi = proj_imp.getRoi()
centre = (roi.getStatistics().xCentroid, roi.getStatistics().yCentroid)
centres.append(centre)
ring_roi_xs = []
ring_roi_ys = []
for theta in range(360):
pt1 = (centre[0] + min_r_pix * math.cos(math.radians(theta)),
centre[1] + min_r_pix * math.sin(math.radians(theta)))
pt2 = (centre[0] + max_r_pix * math.cos(math.radians(theta)),
centre[1] + max_r_pix * math.sin(math.radians(theta)))
roi = Line(pt1[0], pt1[1], pt2[0], pt2[1])
proj_imp.setRoi(roi)
profile = roi.getPixels()
projected_im_row.append(max(profile))
try:
ring_roi_xs.append(roi.getContainedPoints()[profile.index(
max(profile))].x)
except IndexError:
ring_roi_xs.append(pt2[0])
try:
ring_roi_ys.append(roi.getContainedPoints()[profile.index(
max(profile))].y)
except IndexError:
ring_roi_ys.append(pt2[1])
proj_imp.killRoi()
ring_roi = PolygonRoi(ring_roi_xs, ring_roi_ys, Roi.FREELINE)
ring_rois.append(ring_roi)
if generate_roi_stack:
roi_stack.setZ(zidx + 1)
roi_stack.setRoi(ring_roi)
IJ.run(roi_stack, "Line to Area", "")
IJ.run(
roi_stack, "Set...",
"value=" + str(roi_stack.getProcessor().maxValue()) + " slice")
#egfp_imp.setRoi(ring_roi);
projected_im_pix.append(projected_im_row)
# for ch in split_chs:
# ch.close();
out_imp = ImagePlus(
"projected", FloatProcessor([list(x) for x in zip(*projected_im_pix)]))
if generate_roi_stack:
roi_stack.show()
egfp_imp_disp.show()
# merge?
else:
roi_stack = None
return out_imp, roi_stack, ring_rois, centres
def analyze(iDataSet, tbModel, p, output_folder):
#
# LOAD FILES
#
filepath = tbModel.getFileAPth(iDataSet, "RAW", "IMG")
filename = tbModel.getFileName(iDataSet, "RAW", "IMG")
print("Analyzing: "+filepath)
IJ.run("Bio-Formats Importer", "open=["+filepath+"] color_mode=Default view=Hyperstack stack_order=XYCZT");
imp = IJ.getImage()
#
# INIT
#
IJ.run("Options...", "iterations=1 count=1");
#
# SCALING
#
IJ.run(imp, "Scale...", "x="+str(p["scale"])+" y="+str(p["scale"])+" z=1.0 interpolation=Bilinear average process create");
imp = IJ.getImage()
# save output file
output_file = filename+"--downscale_input.tif"
IJ.saveAs(IJ.getImage(), "TIFF", os.path.join(output_folder, output_file))
tbModel.setFileAPth(output_folder, output_file, iDataSet, "INPUT","IMG")
#
# CONVERSION
#
#IJ.run(imp, "8-bit", "");
#
# CROPPING
#
#imp.setRoi(392,386,750,762);
#IJ.run(imp, "Crop", "");
#
# BACKGROUND SUBTRACTION
#
# IJ.run(imp, "Subtract...", "value=32768 stack");
IJ.run(imp, "Z Project...", "projection=[Average Intensity]");
imp_avg = IJ.getImage()
ic = ImageCalculator();
imp = ic.run("Subtract create 32-bit stack", imp, imp_avg);
#
# REGION SEGMENTATION
#
imp1 = Duplicator().run(imp, 1, imp.getImageStackSize()-1)
imp2 = Duplicator().run(imp, 2, imp.getImageStackSize())
imp_diff = ic.run("Subtract create 32-bit stack", imp1, imp2);
#imp_diff.show()
IJ.run(imp_diff, "Z Project...", "projection=[Standard Deviation]");
imp_diff_sd = IJ.getImage()
# save
IJ.run(imp_diff_sd, "Gaussian Blur...", "sigma=5");
output_file = filename+"--sd.tif"
IJ.saveAs(imp_diff_sd, "TIFF", os.path.join(output_folder, output_file))
tbModel.setFileAPth(output_folder, output_file, iDataSet, "SD","IMG")
IJ.run(imp_diff_sd, "Enhance Contrast", "saturated=0.35");
IJ.run(imp_diff_sd, "8-bit", "");
IJ.run(imp_diff_sd, "Properties...", "unit=p pixel_width=1 pixel_height=1 voxel_depth=1");
IJ.run(imp_diff_sd, "Auto Local Threshold", "method=Niblack radius=60 parameter_1=2 parameter_2=0 white");
rm = ROIManipulator.getEmptyRm()
IJ.run(imp_diff_sd, "Analyze Particles...", "add");
# select N largest Rois
diameter_roi = []
for i in range(rm.getCount()):
roi = rm.getRoi(i)
diameter_roi.append([roi.getFeretsDiameter(), roi])
diameter_roi = sorted(diameter_roi, reverse=True)
#print diameter_roi
rm.reset()
for i in range(min(len(diameter_roi), p["n_rois"])):
rm.addRoi(diameter_roi[i][1])
# save
output_file = filename+"--rois"
ROIManipulator.svRoisToFl(output_folder, output_file, rm.getRoisAsArray())
tbModel.setFileAPth(output_folder, output_file+".zip", iDataSet, "REGIONS","ROI")
#
# FFT in each region
#
IJ.run(imp, "Variance...", "radius=2 stack");
output_file = filename+"--beats.tif"
IJ.saveAs(imp, "TIFF", os.path.join(output_folder, output_file))
tbModel.setFileAPth(output_folder, output_file, iDataSet, "BEATS","IMG")
n = rm.getCount()
for i_roi in range(n):
imp_selection = Duplicator().run(imp)
rm.select(imp_selection, i_roi)
IJ.run(imp_selection, "Clear Outside", "stack");
imp_selection.show()
# FFT using Parallel FFTJ
transformer = FloatTransformer(imp_selection.getStack())
transformer.fft()
imp_fft = transformer.toImagePlus(SpectrumType.FREQUENCY_SPECTRUM)
imp_fft.show()
# Analyze FFt
IJ.run(imp_fft, "Gaussian Blur 3D...", "x=0 y=0 z=1.5");
IJ.run(imp_fft, "Plot Z-axis Profile", "");
output_file = filename+"--Region"+str(i_roi+1)+"--fft.tif"
IJ.saveAs(IJ.getImage(), "TIFF", os.path.join(output_folder, output_file))
tbModel.setFileAPth(output_folder, output_file, iDataSet, "FFT_R"+str(i_roi+1),"IMG")
IJ.run(imp_fft, "Select All", "");
rm.addRoi(imp_fft.getRoi())
rm.select(rm.getCount())
rt = ResultsTable()
rt = rm.multiMeasure(imp_fft); #print(rt.getColumnHeadings);
x = rt.getColumn(rt.getColumnIndex("Mean1"))
#rm.runCommand("delete")
peak_height_pos = []
x_min = 10
for i in range(x_min,len(x)/2):
before = x[i-1]
center = x[i]
after = x[i+1]
if (center>before) and (center>after):
peak_height_pos.append([float(x[i]),i])
if len(peak_height_pos)>0:
peak_height_pos = sorted(peak_height_pos, reverse=True)
n_max = 3
for i_max in range(min(len(peak_height_pos),n_max)):
tbModel.setNumVal(round(float(len(x))/float(peak_height_pos[i_max][1]),2), iDataSet, "F"+str(i_max+1)+"_R"+str(i_roi+1))
tbModel.setNumVal(int(peak_height_pos[i_max][0]), iDataSet, "A"+str(i_max+1)+"_R"+str(i_roi+1))
reload(Utility) import CountParticles reload(CountParticles) from CountParticles import countParticles #inputDirectory='/home/bnorthan/Brian2012/Round2/RogueImageJPlugins/SpotDetection/Images/' #inputName='001-D0_cropped.tif' #dataset=data.open(inputDirectory+inputName) #display.createDisplay(dataset.getName(), dataset) inputImp=IJ.getImage() inputDataset=Utility.getDatasetByName(data, inputImp.getTitle()) truecolor1=Duplicator().run(inputImp) truecolor1.show() # get the roi that will be processed inputRoi=inputImp.getRoi().clone() inputRec = inputRoi.getBounds() x1=long(inputRec.getX()) y1=long(inputRec.getY()) x2=x1+long(inputRec.getWidth())-1 y2=y1+long(inputRec.getHeight())-1 # crop the roi interval=FinalInterval( array([x1, y1 ,0], 'l'), array([x2, y2, 2], 'l') ) cropped=ops.crop(interval, None, inputDataset.getImgPlus() ) dataset=data.create(cropped)
########## Use thresholding and selection to define UFOV ###################################################################################
#IJ.run("ROI Manager...", "") # not sure if I need this
IJ.setRawThreshold(
original, 1, 255,
'') # background pixels have value 0. See IMAGE>ADJUST>THRESHOLD
IJ.run(original, "Create Selection",
"") # add bounding box. See EDIT>SELECTION
IJ.run(original, "To Bounding Box",
"") # this box defines the UFOV. See EDIT>SELECTION
IJ.resetThreshold(
original) # get back original now UFOV is definedthresholding
UFOV = Duplicator().run(
original) # duplicate the original image, only the CFOV
UFOV.setTitle("UFOV")
UFOV.show()
CFOV_fraction = 0.75 # choose the fraction of the UFOV that defines the CFOV
IJ.run(original, "Scale... ", "x=" + str(CFOV_fraction) + " y=" +
str(CFOV_fraction) + " centered") # rescale bounding box to get CFOV
CFOV = Duplicator().run(
original) # duplicate the original image, only the CFOV
CFOV.setTitle("CFOV")
CFOV.show()
######### Nema process including Re-bin image to larger pixels ################################################################################
desired_pixel_width = getPixel(
) # 6.4 mm default, remember tolerance is +/-30%
current_pixel_width = CFOV.getCalibration(
).pixelWidth #get pixel width, 1.16 mm
# ROI CREATION
image_mip.show()
image_mip.setRoi(image_mip.width / 2, image_mip.height / 2, regionSize,
regionSize)
from ij.gui import WaitForUserDialog
myWait = WaitForUserDialog("Choose ROI",
"Please position the ROI.. Press 'OK' when done.")
myWait.show()
# CROP
image.setRoi(image_mip.getRoi())
from ij import IJ
image = Duplicator().run(image)
image.show()
image_mip.close()
image = ImageJFunctions.wrap(image)
#
def roi(mask, image):
# Convert ROI from R^n to Z^n.
#discreteROI = Views.raster(Masks.toRealRandomAccessible(mask))
# Apply finite bounds to the discrete ROI.
boundedDiscreteROI = Views.interval(mask, image)
# Create an iterable version of the finite discrete ROI.
iterableROI = Regions.iterable(boundedDiscreteROI)
return Regions.sample(iterableROI, image)
def runOneFile(fullFilePath):
global gNumChannels
global gAlignBatchVersion
if not os.path.isfile(fullFilePath):
bPrintLog(
'\nERROR: runOneFile() did not find file: ' + fullFilePath + '\n',
0)
return 0
bPrintLog(
time.strftime("%H:%M:%S") + ' starting runOneFile(): ' + fullFilePath,
1)
enclosingPath = os.path.dirname(fullFilePath)
head, tail = os.path.split(enclosingPath)
enclosingPath += '/'
#make output folders
destFolder = enclosingPath + tail + '_channels/'
if not os.path.isdir(destFolder):
os.makedirs(destFolder)
destMaxFolder = destFolder + 'max/'
if not os.path.isdir(destMaxFolder):
os.makedirs(destMaxFolder)
if gDoAlign:
destAlignmentFolder = destFolder + 'alignment/'
if not os.path.isdir(destAlignmentFolder):
os.makedirs(destAlignmentFolder)
if gSave8bit:
eightBitFolder = destFolder + 'channels8/'
if not os.path.isdir(eightBitFolder):
os.makedirs(eightBitFolder)
eightBitMaxFolder = eightBitFolder + 'max/'
if not os.path.isdir(eightBitMaxFolder):
os.makedirs(eightBitMaxFolder)
# open image
imp = Opener().openImage(fullFilePath)
# get parameters of image
(width, height, nChannels, nSlices, nFrames) = imp.getDimensions()
bitDepth = imp.getBitDepth()
infoStr = imp.getProperty("Info") #get all .tif tags
if not infoStr:
infoStr = ''
infoStr += 'bAlignBatch_Version=' + str(gAlignBatchVersion) + '\n'
infoStr += 'bAlignBatch_Time=' + time.strftime(
"%Y%m%d") + '_' + time.strftime("%H%M%S") + '\n'
msgStr = 'w:' + str(width) + ' h:' + str(height) + ' slices:' + str(nSlices) \
+ ' channels:' + str(nChannels) + ' frames:' + str(nFrames) + ' bitDepth:' + str(bitDepth)
bPrintLog(msgStr, 1)
path, filename = os.path.split(fullFilePath)
shortName, fileExtension = os.path.splitext(filename)
#
# look for num channels in ScanImage infoStr
if gGetNumChanFromScanImage:
for line in infoStr.split('\n'):
#scanimage.SI4.channelsSave = [1;2]
scanimage4 = find(line, 'scanimage.SI4.channelsSave =') == 0
#state.acq.numberOfChannelsSave=2
scanimage3 = find(line, 'state.acq.numberOfChannelsSave=') == 0
if scanimage3:
#print 'line:', line
equalIdx = find(line, '=')
line2 = line[equalIdx + 1:]
if gGetNumChanFromScanImage:
gNumChannels = int(line2)
bPrintLog(
'over-riding gNumChannels with: ' + str(gNumChannels),
2)
if scanimage4:
#print ' we have a scanimage 4 file ... now i need to exptract the number of channel'
#print 'line:', line
equalIdx = find(line, '=')
line2 = line[equalIdx + 1:]
for delim in ';[]':
line2 = line2.replace(delim, ' ')
if gGetNumChanFromScanImage:
gNumChannels = len(line2.split())
bPrintLog(
'over-riding gNumChannels with: ' + str(gNumChannels),
2)
# show
imp.show()
# split channels if necc. and grab the original window names
if gNumChannels == 1:
origImpWinStr = imp.getTitle() #use this when only one channel
origImpWin = WindowManager.getWindow(
origImpWinStr) #returns java.awt.Window
if gNumChannels == 2:
winTitle = imp.getTitle()
bPrintLog('Deinterleaving 2 channels...', 1)
IJ.run('Deinterleave',
'how=2 keep') #makes ' #1' and ' #2', with ' #2' frontmost
origCh1WinStr = winTitle + ' #1'
origCh2WinStr = winTitle + ' #2'
origCh1Imp = WindowManager.getImage(origCh1WinStr)
origCh2Imp = WindowManager.getImage(origCh2WinStr)
origCh1File = destFolder + shortName + '_ch1.tif'
origCh2File = destFolder + shortName + '_ch2.tif'
# work on a copy, mostly for alignment with cropping
copy = Duplicator().run(imp)
#copy.copyAttributes(imp) #don't copy attributes, it copies the name (which we do not want)
copy.show()
#
# crop (on copy)
if gDoCrop:
bPrintLog('making cropping rectangle (left,top,width,height) ', 1)
bPrintLog(
str(gCropLeft) + ' ' + str(gCropTop) + ' ' + str(gCropWidth) +
' ' + str(gCropHeight), 2)
roi = Roi(gCropLeft, gCropTop, gCropWidth,
gCropHeight) #left,top,width,height
copy.setRoi(roi)
time.sleep(
0.5
) # otherwise, crop SOMETIMES failes. WHAT THE F**K FIJI DEVELOPERS, REALLY, WHAT THE F**K
#bPrintLog('cropping', 1)
IJ.run('Crop')
infoStr += 'bCropping=' + str(gCropLeft) + ',' + str(
gCropTop) + ',' + str(gCropWidth) + ',' + str(gCropHeight) + '\n'
#
# remove calibration ( on original)
if gRemoveCalibration:
cal = imp.getCalibration()
calCoeff = cal.getCoefficients()
if calCoeff:
msgStr = 'Calibration is y=a+bx' + ' a=' + str(
calCoeff[0]) + ' b=' + str(calCoeff[1])
bPrintLog(msgStr, 1)
#remove calibration
bPrintLog('\tRemoving Calibration', 2)
imp.setCalibration(None)
#without these, 8-bit conversion goes to all 0 !!! what the f**k !!!
#bPrintLog('calling imp.resetStack() and imp.resetDisplayRange()', 2)
imp.resetStack()
imp.resetDisplayRange()
#get and print out min/max
origMin = StackStatistics(imp).min
origMax = StackStatistics(imp).max
msgStr = '\torig min=' + str(origMin) + ' max=' + str(origMax)
bPrintLog(msgStr, 2)
# 20150723, 'shift everybody over by linear calibration intercept calCoeff[0] - (magic number)
if 1:
# [1] was this
#msgStr = 'Subtracting original min '+str(origMin) + ' from stack.'
#bPrintLog(msgStr, 2)
#subArgVal = 'value=%s stack' % (origMin,)
#IJ.run('Subtract...', subArgVal)
# [2] now this
#msgStr = 'Adding calCoeff[0] '+str(calCoeff[0]) + ' from stack.'
#bPrintLog(msgStr, 2)
#addArgVal = 'value=%s stack' % (int(calCoeff[0]),)
#IJ.run('Add...', addArgVal)
# [3] subtract a magic number 2^15-2^7 = 32768 - 128
magicNumber = gLinearShift #2^15 - 128
msgStr = 'Subtracting a magic number (linear shift) ' + str(
magicNumber) + ' from stack.'
bPrintLog(msgStr, 2)
infoStr += 'bLinearShift=' + str(gLinearShift) + '\n'
subArgVal = 'value=%s stack' % (gLinearShift, )
IJ.run(imp, 'Subtract...', subArgVal)
# 20150701, set any pixel <0 to 0
if 0:
ip = imp.getProcessor() # returns a reference
pixels = ip.getPixels() # returns a reference
msgStr = '\tSet all pixels <0 to 0. This was added 20150701 ...'
bPrintLog(msgStr, 2)
pixels = map(lambda x: 0 if x < 0 else x, pixels)
bPrintLog('\t\t... done', 2)
#get and print out min/max
newMin = StackStatistics(imp).min
newMax = StackStatistics(imp).max
msgStr = '\tnew min=' + str(newMin) + ' max=' + str(newMax)
bPrintLog(msgStr, 2)
#append calibration to info string
infoStr += 'bCalibCoeff_a = ' + str(calCoeff[0]) + '\n'
infoStr += 'bCalibCoeff_b = ' + str(calCoeff[1]) + '\n'
infoStr += 'bNewMin = ' + str(newMin) + '\n'
infoStr += 'bNewMax = ' + str(newMax) + '\n'
#
# set up
if gNumChannels == 1:
impWinStr = copy.getTitle() #use this when only one channel
impWin = WindowManager.getWindow(impWinStr) #returns java.awt.Window
if gNumChannels == 2:
winTitle = copy.getTitle()
bPrintLog('Deinterleaving 2 channels...', 1)
IJ.run('Deinterleave',
'how=2 keep') #makes ' #1' and ' #2', with ' #2' frontmost
ch1WinStr = winTitle + ' #1'
ch2WinStr = winTitle + ' #2'
ch1Imp = WindowManager.getImage(ch1WinStr)
ch2Imp = WindowManager.getImage(ch2WinStr)
ch1File = destFolder + shortName + '_ch1.tif'
ch2File = destFolder + shortName + '_ch2.tif'
#
# alignment
if gDoAlign and gNumChannels == 1 and copy.getNSlices() > 1:
infoStr += 'AlignOnChannel=1' + '\n'
#snap to middle slice
if gAlignOnMiddleSlice:
middleSlice = int(
math.floor(copy.getNSlices() /
2)) #int() is necc., python is f*****g picky
else:
middleSlice = gAlignOnThisSlice
copy.setSlice(middleSlice)
transformationFile = destAlignmentFolder + shortName + '.txt'
bPrintLog('MultiStackReg aligning:' + impWinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=Align file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body] save' % (
impWin, transformationFile)
IJ.run('MultiStackReg', stackRegParams)
infoStr += 'AlignOnSlice=' + str(middleSlice) + '\n'
#20150723, we just aligned on a cropped copy, apply alignment to original imp
origImpTitle = imp.getTitle()
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' % (
origImpTitle, transformationFile)
IJ.run('MultiStackReg', stackRegParams)
if gDoAlign and gNumChannels == 2 and ch1Imp.getNSlices(
) > 1 and ch2Imp.getNSlices() > 1:
#apply to gAlignThisChannel
alignThisWindow = ''
applyAlignmentToThisWindow = ''
if gAlignThisChannel == 1:
infoStr += 'AlignOnChannel=1' + '\n'
transformationFile = destAlignmentFolder + shortName + '_ch1.txt'
alignThisWindow = ch1WinStr
applyAlignmentToThisWindow = ch2WinStr
else:
infoStr += 'AlignOnChannel=2' + '\n'
transformationFile = destAlignmentFolder + shortName + '_ch2.txt'
alignThisWindow = ch2WinStr
applyAlignmentToThisWindow = ch1WinStr
alignThisImp = WindowManager.getImage(alignThisWindow)
#snap to middle slice
if gAlignOnMiddleSlice:
middleSlice = int(
math.floor(alignThisImp.getNSlices() /
2)) #int() is necc., python is f*****g picky
else:
middleSlice = gAlignOnThisSlice
alignThisImp.setSlice(middleSlice)
infoStr += 'bAlignOnSlice=' + str(middleSlice) + '\n'
bPrintLog('MultiStackReg aligning:' + alignThisWindow, 1)
stackRegParams = 'stack_1=[%s] action_1=Align file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body] save' % (
alignThisWindow, transformationFile)
IJ.run('MultiStackReg', stackRegParams)
# 20150723, we just aligned on a copy, apply alignment to both channels of original
# ch1
bPrintLog('MultiStackReg applying alignment to:' + origCh1WinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' % (
origCh1WinStr, transformationFile)
IJ.run('MultiStackReg', stackRegParams)
# ch2
bPrintLog('MultiStackReg applying alignment to:' + origCh2WinStr, 1)
stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' % (
origCh2WinStr, transformationFile)
IJ.run('MultiStackReg', stackRegParams)
#apply alignment to other window
#bPrintLog('MultiStackReg applying alignment to:' + applyAlignmentToThisWindow, 1)
#applyAlignThisImp = WindowManager.getImage(applyAlignmentToThisWindow)
#stackRegParams = 'stack_1=[%s] action_1=[Load Transformation File] file_1=[%s] stack_2=None action_2=Ignore file_2=[] transformation=[Rigid Body]' %(applyAlignmentToThisWindow,transformationFile)
#IJ.run('MultiStackReg', stackRegParams)
elif gDoAlign:
bPrintLog('Skipping alignment, there may be only one slice?', 3)
#
# save
if gNumChannels == 1:
imp.setProperty("Info", infoStr)
impFile = destFolder + shortName + '.tif'
#bPrintLog('Saving:' + impFile, 1)
bSaveStack(imp, impFile)
#max project
bSaveZProject(imp, destMaxFolder, shortName)
if gNumChannels == 2:
#ch1
origCh1Imp.setProperty("Info", infoStr)
#bPrintLog('Saving:' + ch1File, 1)
bSaveStack(origCh1Imp, ch1File)
#max project
bSaveZProject(origCh1Imp, destMaxFolder, shortName + '_ch1')
#ch2
origCh2Imp.setProperty("Info", infoStr)
#bPrintLog('Saving:' + ch2File, 1)
bSaveStack(origCh2Imp, ch2File)
#max project
bSaveZProject(origCh2Imp, destMaxFolder, shortName + '_ch2')
#
# post convert to 8-bit and save
if gSave8bit:
if bitDepth == 16:
if gNumChannels == 1:
bPrintLog('Converting to 8-bit:' + impWinStr, 1)
IJ.selectWindow(impWinStr)
#IJ.run('resetMinAndMax()')
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(imp, impFile)
#max project
bSaveZProject(imp, eightBitMaxFolder, shortName)
if gNumChannels == 2:
#
bPrintLog('Converting to 8-bit:' + origCh1WinStr, 1)
IJ.selectWindow(origCh1WinStr)
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch1.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(origCh1Imp, impFile)
#max project
bSaveZProject(origCh1Imp, eightBitMaxFolder,
shortName + '_ch1')
#
bPrintLog('Converting to 8-bit:' + origCh2WinStr, 1)
IJ.selectWindow(origCh2WinStr)
#IJ.run('resetMinAndMax()')
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch2.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(origCh2Imp, impFile)
#max project
bSaveZProject(origCh2Imp, eightBitMaxFolder,
shortName + '_ch2')
#
# close original window
imp.changes = 0
imp.close()
#copy
copy.changes = 0
copy.close()
#
# close ch1/ch2
if gNumChannels == 2:
#original
origCh1Imp.changes = 0
origCh1Imp.close()
origCh2Imp.changes = 0
origCh2Imp.close()
#copy
ch1Imp.changes = 0
ch1Imp.close()
ch2Imp.changes = 0
ch2Imp.close()
bPrintLog(
time.strftime("%H:%M:%S") + ' finished runOneFile(): ' + fullFilePath,
1)
def main():
#print (sys.version_info) # debug
#print(sys.path) # debug
data_root = r'C:\Users\dougk\Desktop\test'
# debug
output_root = r'C:\Users\dougk\Desktop\test'
#debug
#default_directory = r'C:\\Users\\Doug\\Desktop\\test';
#data_root, output_root = file_location_chooser(default_directory);
if (data_root is None) or (output_root is None):
raise IOError("File location dialogs cancelled!")
timestamp = datetime.strftime(datetime.now(), "%Y-%m-%d %H.%M.%S")
output_path = os.path.join(output_root, (timestamp + " output"))
for file_path in filterByFileType(os.listdir(data_root), '.tif'):
subfolder_name = os.path.splitext(file_path)[0]
output_subfolder = os.path.join(output_path, subfolder_name)
print(output_subfolder)
os.makedirs(output_subfolder)
imps = bf.openImagePlus(os.path.join(data_root, file_path))
imp = imps[0]
imp.show()
h = imp.height
w = imp.width
slices = imp.getNSlices()
channels = imp.getNChannels()
frames = imp.getNFrames()
# rotation step - since using multiples of 90, TransformJ.Turn is more efficient
IJ.run("Enhance Contrast", "saturated=0.35")
angleZ = 1
while ((angleZ % 90) > 0):
gd = GenericDialog("Rotate?")
gd.addMessage(
"Define rotation angle - increments of 90. Apical at top")
gd.addNumericField("Rotation angle", 0, 0)
gd.showDialog()
angleZ = int(gd.getNextNumber())
if (angleZ > 1):
IJ.run("TransformJ Turn",
"z-angle=" + str(angleZ) + " y-angle=0 x-angle=0")
imp.close()
imp = WindowManager.getCurrentImage()
imp.setTitle(file_path)
# trim time series
IJ.run("Enhance Contrast", "saturated=0.35")
imp.setDisplayMode(IJ.COLOR)
WaitForUserDialog(
"Scroll to the first frame of the period of interest and click OK"
).show()
start_frame = imp.getT()
WaitForUserDialog(
"Scroll to the last frame of the period of interest and click OK"
).show()
end_frame = imp.getT()
trim_imp = Duplicator().run(imp, 1, channels, 1, slices, start_frame,
end_frame)
imp.close()
trim_imp.show()
dup_imp = Duplicator().run(trim_imp)
# create images to process and find bounds for
dup_imps = ChannelSplitter().split(dup_imp)
myo_imp = dup_imps[1]
mem_imp = dup_imps[0]
FileSaver(myo_imp).saveAsTiffStack(
os.path.join(output_subfolder, "myosin_channel.tif"))
FileSaver(mem_imp).saveAsTiffStack(
os.path.join(output_subfolder, "membrane_channel.tif"))
# set basal bounds
myo_imp.show()
ImageConverter(myo_imp).convertToGray8()
frames = myo_imp.getNFrames()
gb = GaussianBlur()
for fridx in range(0, frames):
myo_imp.setSliceWithoutUpdate(fridx + 1)
ip = myo_imp.getProcessor()
gb.blurGaussian(ip, 5.0, 1.0, 0.02)
# assymmetrical Gaussian
IJ.run(myo_imp, "Convert to Mask",
"method=Otsu background=Dark calculate")
IJ.run("Despeckle", "stack")
title = myo_imp.getTitle()
# assume that first frame is good quality image...
basal_edges = find_basal_edges(myo_imp)
#myo_imp.hide()
mem_imp.hide()
# draw some edges for checking
roim = RoiManager()
xs = [x for x in range(1,
trim_imp.getWidth() + 1)]
trim_imp.show()
for fridx in range(0, myo_imp.getNFrames()):
trim_imp.setPosition(2, 1, fridx + 1)
IJ.run("Enhance Contrast", "saturated=0.35")
roi = PolygonRoi(xs, basal_edges[fridx], Roi.POLYLINE)
trim_imp.setRoi(roi)
roim.addRoi(roi)
# apply a threshold to detect objects (cells, nuclei,...)
IJ.setAutoThreshold(imp, "Otsu")
Prefs.blackBackground = False
IJ.run(imp, "Convert to Mask", "")
# analyse objects and put results in table
IJ.run("Set Measurements...", "area display redirect=None decimal=5")
IJ.run(imp, "Analyze Particles...", "exclude add")
manager = RoiManager.getRoiManager()
manager.runCommand("Save",
file.toString() + ".RoiSet.zip")
for roi in manager.getRoisAsArray():
copy.setRoi(roi)
copy.show()
IJ.run(copy, "Enlarge...", "enlarge=-2")
# analyse the image
statistics = copy.getStatistics()
print("mean: " + str(statistics.mean))
print("area:" + str(statistics.area))
# write the analysed valurs in a table
table = ResultsTable.getResultsTable()
table.incrementCounter()
table.addValue("filename", file.toString())
table.addValue("mean_robert", statistics.mean)
table.addValue("area", statistics.area)
table.addValue("total instensity",
statistics.mean * statistics.area)
reload(Utility) import CountParticles reload(CountParticles) from CountParticles import countParticles #inputDirectory='/home/bnorthan/Brian2012/Round2/RogueImageJPlugins/SpotDetection/Images/' #inputName='001-D0_cropped.tif' #dataset=data.open(inputDirectory+inputName) #display.createDisplay(dataset.getName(), dataset) inputImp = IJ.getImage() inputDataset = Utility.getDatasetByName(data, inputImp.getTitle()) truecolor1 = Duplicator().run(inputImp) truecolor1.show() # get the roi that will be processed inputRoi = inputImp.getRoi().clone() inputRec = inputRoi.getBounds() x1 = long(inputRec.getX()) y1 = long(inputRec.getY()) x2 = x1 + long(inputRec.getWidth()) - 1 y2 = y1 + long(inputRec.getHeight()) - 1 # crop the roi interval = FinalInterval(array([x1, y1, 0], 'l'), array([x2, y2, 2], 'l')) cropped = ops.crop(interval, None, inputDataset.getImgPlus()) dataset = data.create(cropped)
#print(centres);
for ch in split_chs:
ch.close()
w = len(projected_im_row)
h = len(projected_im_pix)
ip = FloatProcessor(w, h)
pix = ip.getPixels()
for x in range(w):
for y in range(h):
pix[y * w + x] = projected_im_pix[y][x]
out_imp = ImagePlus("projected", ip)
out_imp.show()
egfp_imp_disp.show()
roi_stack.show()
#egfp_imp_disp.addImageListener(UpdateRoiImageListener(ring_rois));
unwrapped_imp = IJ.createImage("twisted unwrap", 2 * out_imp.getWidth(),
out_imp.getHeight(), 1, 32)
unwrapped_imp.show()
IJ.setTool("freeline")
WaitForUserDialog("input unwrap axis").show()
unwrap_poly = out_imp.getRoi().getPolygon()
unwrap_poly_xs
#print("unzip axis = " + str(unzip_axis));
#right_side_xs = unzip_axis;
#right_side_xs.append(out_imp.getWidth());
#right_side_xs.append(out_imp.getWidth());
str(metadata['x_unit']) + " pixel_width=" +
str(metadata['x_physical_size']) + " pixel_height=" +
str(metadata['y_physical_size']) + " voxel_depth=" +
str(metadata['z_extent'] / metadata['z_pixels']))
imp.show()
imp, z_lims = z_crop(imp)
use_ch = imp.getC()
print(use_ch)
IJ.setTool("rect")
imp.hide()
imp2 = ZProjector.run(imp, "max")
imp2.show()
imp2.setC(use_ch + 1)
WaitForUserDialog("Select XY region to crop...").show()
crop_roi = imp2.getRoi()
imp2.close()
imp.show()
imp.setRoi(crop_roi)
IJ.run("Crop", "")
frames = imp.getNFrames()
slices = imp.getNSlices()
imp = Duplicator().run(imp, use_ch + 1, use_ch + 1, 1, slices, 1, frames)
imp.show()
# now do 3d segmentation....
#histo = StackStatistics(imp).histogram;
#global_thr = AutoThresholder().getThreshold(AutoThresholder.Method.IJ_IsoData, histo) + 1;
#print(global_thr);
#IJ.run(imp, "3D Simple Segmentation", "low_threshold=" + str(global_thr) + " min_size=200 max_size=-1");
#segimp = WM.getImage("Seg");
