def process_pi_signal(path, position, unsynchronized=True):
if unsynchronized:
path_signal = path + "\\pi"
path_signal_before = path_signal + "\\before"
path_signal_after = path_signal + "\\after"
path_signal_merged = path_signal + "\\merged"
path_imp_before = path_signal_before + "\\pi_in-focusxy%sc1.tif" % position
path_imp_after = path_signal_after + "\\pixy%sc1.tif" % position
path_imp_merged = path_signal_merged + "\\merged.tif"
path_imp_merged_sub = path_signal_merged + "\\merged_sub.tif"
imp1 = IJ.openImage(path_imp_before)
imp1.show()
imp2 = IJ.openImage(path_imp_after)
imp2.show()
zp1 = ZProjector(imp1)
zp1.setMethod(ZProjector.AVG_METHOD)
zp1.doProjection()
zpimp1 = zp1.getProjection()
zp2 = ZProjector(imp2)
zp2.setMethod(ZProjector.AVG_METHOD)
zp2.doProjection()
zpimp2 = zp2.getProjection()
imp_sub1 = ImageCalculator().run("Subtract create stack", imp1, zpimp1)
imp_sub1.show()
imp_sub2 = ImageCalculator().run("Subtract create stack", imp2, zpimp2)
imp_sub2.show()
concatenate_files(imp1, imp2, path_imp_merged)
concatenate_files(imp_sub1, imp_sub2, path_imp_merged_sub)
else:
path_signal = path + "\\pi\\seq0002xy%sc1.tif" % position
path_sub = path + "\\pi\\sub.tif"
imp = IJ.openImage(path_signal)
imp.show()
zp = ZProjector(imp)
zp.setMethod(ZProjector.AVG_METHOD)
zp.doProjection()
zpimp = zp.getProjection()
imp_sub = ImageCalculator().run("Subtract create stack", imp, zpimp)
imp_sub.show()
IJ.saveAs(imp_sub, "Tiff", path_sub)
imp.changes = False
imp.close()
zpimp.changes = False
zpimp.close()
imp_sub.changes = False
imp_sub.close()
def maxZprojection(stackimp):
'''copied from EMBL CMCI python-ImageJ cookbook.'''
zp = ZProjector(stackimp)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
zpimp = zp.getProjection()
return zpimp
def getMIP(stackImp):
"""
Create a maximum intensity projection (MIP) from an ImagePlus stack
@return ImagePlus maximum projection
"""
zp = ZProjector(stackImp)
zp.setMethod(ZProjector.MAX_METHOD)
zp.setStopSlice(stackImp.getNSlices())
zp.doHyperStackProjection(False)
return zp.getProjection()
def saveMaxProject(self, destFolder=''):
# ch1
if self.imp_ch1:
# make max project
zp = ZProjector(self.imp_ch1)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
zimp = zp.getProjection()
# save
fs = FileSaver(zimp)
bPrintLog('saveMaxProject():' + self.savePathMax_ch1, 3)
fs.saveAsTiff(self.savePathMax_ch1)
# ch2
if self.imp_ch2:
# make max project
zp = ZProjector(self.imp_ch2)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
zimp = zp.getProjection()
# save
fs = FileSaver(zimp)
bPrintLog('saveMaxProject():' + self.savePathMax_ch2, 3)
fs.saveAsTiff(self.savePathMax_ch2)
# ch1
if self.imp_ch3:
# make max project
zp = ZProjector(self.imp_ch3)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
zimp = zp.getProjection()
# save
fs = FileSaver(zimp)
bPrintLog('saveMaxProject():' + self.savePathMax_ch3, 3)
fs.saveAsTiff(self.savePathMax_ch3)
def projectionImage(imp):
"""Returns the MIP of the specified ImagePlus (a composite stack)"""
from ij.plugin import ZProjector
roi_exists = imp.getRoi() is not None
imp.deleteRoi()
zp = ZProjector(imp)
zp.setMethod(ZProjector.MAX_METHOD)
zp.setStartSlice(1)
zp.setStopSlice(imp.getNSlices())
zp.doHyperStackProjection(True)
mip_imp = zp.getProjection()
mip_imp.setCalibration(imp.getCalibration())
if roi_exists:
mip_imp.restoreRoi()
return mip_imp
def project_z(imp, method):
zp = ZProjector(imp)
if method=="max":
zp.setMethod(ZProjector.MAX_METHOD)
if method=="sum":
zp.setMethod(ZProjector.SUM_METHOD)
zp.setStopSlice(imp.getNSlices())
zp.doHyperStackProjection(True)
zpimp = zp.getProjection()
#IJ.run(imp, "Z Project...", "projection=[Max Intensity] all");
#impout = IJ.getImage()
return zpimp
def Zproj(stackimp, method, z_ind, z_range):
# Gets a stack and max project it.
# imp= stackimp.duplicate()
zp = ZProjector(stackimp)
if method == "MAX":
print "MAX"
zp.setMethod(ZProjector.MAX_METHOD)
elif method == "SUM":
print "SUM"
zp.setMethod(ZProjector.MAX_METHOD)
print "+/-", int(z_range / 2), "of z-index", z_ind
zp.setStartSlice(z_ind - int(z_range / 2))
zp.setStopSlice(z_ind + int(z_range / 2))
zp.doProjection()
zpimp = zp.getProjection()
return zpimp
def BackgroundFilter(imp, projection_method = "Median"):
title = imp.getTitle()
#Make a dict containg method_name:const_fieled_value pairs for the projection methods
methods_as_strings=['Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices', 'Standard Deviation', 'Median']
methods_as_const=[ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD, ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD]
method_dict=dict(zip(methods_as_strings, methods_as_const))
#The Z-Projection magic happens here through a ZProjector object
zp = ZProjector(imp)
zp.setMethod(method_dict[projection_method])
zp.doProjection()
outstack = imp.createEmptyStack()
outstack.addSlice(zp.getProjection().getProcessor())
imp2 = ImagePlus(title+'_'+projection_method, outstack)
out = ImageCalculator().run("Subtract create 32-bit stack", imp, imp2)
return out
def process_caspase_signal(path_signal, path_imp, path_imp_out):
path_imp = path_signal + path_imp
imp = IJ.openImage(path_imp)
imp.show()
zp = ZProjector(imp)
zp.setMethod(ZProjector.AVG_METHOD)
zp.doProjection()
zpimp = zp.getProjection()
imp_sub = ImageCalculator().run("Subtract create stack", imp, zpimp)
imp_sub.show()
IJ.saveAs(imp_sub, "Tiff", path_signal + path_imp_out)
imp.changes = False
imp.close()
imp_sub.changes = False
imp_sub.close()
def Zproj(stackimp,method,z_ind,z_range): """ Gets a single channel z-stack as imp and max project it using the provided method. stackimp(ImagePlus): a single channel z-stack as ImagePlus object z_index(int): The central stack number z_range(int): The total number of stacks surrounding the z_index to indclude """ from ij.plugin import ZProjector # imp= stackimp.duplicate() zp = ZProjector(stackimp) if method=="MAX": print "MAX" zp.setMethod(ZProjector.MAX_METHOD) elif method=="SUM": zp.setMethod(ZProjector.MAX_METHOD) zp.setStartSlice(z_ind-int(z_range/2)) zp.setStopSlice(z_ind+int(z_range/2)) zp.doProjection() zpimp = zp.getProjection() return zpimp
def subtractzproject(imp, projectionMethod="Median"):
"""This function takes an input stack, and subtracts a projection from the
whole stack from each individual frame. Thereby, everything that is not moving
in a timeseries is filtered away.
Args:
imp (ImagePlus): An input stack as ImagePlus object.
projectionMethod (str, optional): Choose the projection method. Options are
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices', 'Standard Deviation', 'Median'.
Defaults to "Median".
Returns:
ImagePlus: The resulting stack.
"""
#Start by getting the active image window and get the current active channel and other stats
cal = imp.getCalibration()
title = imp.getTitle()
# Define a dictionary containg method_name:const_fieled_value pairs for the projection methods.
methods_as_strings = [
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices',
'Standard Deviation', 'Median'
]
methods_as_const = [
ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD,
ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD
]
method_dict = dict(zip(methods_as_strings, methods_as_const))
# Run Z-Projection.
zp = ZProjector(imp)
zp.setMethod(method_dict[projectionMethod])
zp.doProjection()
impMedian = zp.getProjection()
# Subtract Z-Projection and return output ImagePlus.
impout = ImageCalculator().run("Subtract create 32-bit stack", imp,
impMedian)
impout.setCalibration(cal)
return impout
def saveMaxProject(self, destFolder=''):
channelNumber = 1
for imp in self.channelImp:
if not destFolder:
destFolder = os.path.join(self.enclosingPath, self.enclosingfolder + '_tif', 'max')
if not os.path.isdir(destFolder):
os.makedirs(destFolder)
# make max project
zp = ZProjector(imp)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
zimp = zp.getProjection()
# save
saveFile = 'max_' + os.path.splitext(self.filename)[0] + '_ch' + str(channelNumber) + '.tif'
savePath = os.path.join(destFolder, saveFile)
fs = FileSaver(zimp)
bPrintLog('saveMaxProject():' + savePath, 3)
fs.saveAsTiff(savePath)
channelNumber += 1
def startTracking(filepath, fdir, ffile, filename):
outpath = fdir
troutpath = fdir + separator + ffile + separator + "tracked" + separator
stackoutpath = fdir + separator + ffile + separator + "stacks" + separator
pixwidth = 0.647
interval_sec = 600 #pr
if "141006" in fdir:
interval_sec = 600
elif "141117" in fdir:
interval_sec = 300
elif "141215" in fdir:
interval_sec = 300
if not os.path.isdir(outpath):
os.mkdir(outpath)
if not os.path.isdir(troutpath):
os.mkdir(troutpath)
if not os.path.isdir(stackoutpath):
os.mkdir(stackoutpath)
print 'filepath: ', filepath #pr
print 'fdir: ', fdir #pr
print 'ffile: ', ffile #pr
IJ.run("Image Sequence...",
"open=" + filepath + " file=" + filename + " sort")
#pr
#IJ.run("Image Sequence...", "open=" + filepath + " file=molm sort"); #pr
imp = WindowManager.getCurrentImage()
imptitle = imp.getTitle()
nframes = imp.getNSlices()
IJ.run(
imp, "Properties...", "channels=1 slices=1 frames=" + str(nframes) +
" unit=inch pixel_width=" + str(pixwidth) + " pixel_height=" +
str(pixwidth) + " voxel_depth=1.0000 frame=[" + str(interval_sec) +
" sec]")
IJ.run(imp, "Duplicate...", "title=" + imptitle + "_dup duplicate")
imp_dup = WindowManager.getImage(imptitle + "_dup")
IJ.run(imp_dup, "Gaussian Blur...", "sigma=50 stack")
ic = ImageCalculator()
imp_corr = ic.run("Divide create 32-bit stack", imp, imp_dup)
imp_corr.setTitle(imptitle + "_corrected")
imp_corr.show()
imp.changes = False
imp.close()
imp_dup.changes = False
imp_dup.close()
IJ.run(imp_corr, "8-bit", "")
IJ.run(
imp_corr, "Normalize Local Contrast",
"block_radius_x=100 block_radius_y=100 standard_deviations=1 stretch stack"
)
IJ.saveAs(imp_corr, "Tiff",
stackoutpath + separator + imptitle + "_corrected.tif")
IJ.run(imp_corr, "Duplicate...", "title=" + imptitle + "_bg duplicate")
imp_bg = WindowManager.getImage(imptitle + "_bg")
#Prefs.blackBackground = True;
IJ.setAutoThreshold(imp_bg, "MinError dark")
IJ.run(imp_bg, "Convert to Mask", "stack")
IJ.run(imp_bg, "Analyze Particles...",
"size=10000-Infinity show=Masks stack")
imp_bg.changes = False
imp_bg.close()
imp_bgmask = WindowManager.getImage("Mask of " + imptitle + "_bg")
ic = ImageCalculator()
imp_corr_wobg = ic.run("Subtract create stack", imp_corr, imp_bgmask)
imp_corr_wobg.setTitle(imptitle + "_corrected_womask")
imp_corr_wobg.show()
IJ.saveAs(imp_corr_wobg, "Tiff",
stackoutpath + separator + imptitle + "_corrected_womask.tif")
#pr
# pr: substract average frames
zp = ZProjector(imp_corr_wobg)
zp.setMethod(ZProjector.AVG_METHOD)
zp.doProjection()
zpimp = zp.getProjection()
zpimp.show()
imp_corr_wobg_sub = ImageCalculator().run("Subtract create stack",
imp_corr_wobg, zpimp)
imp_corr_wobg_sub.show()
#imp_corr_wobg.changes = False
#imp_corr_wobg.close()
# pr: subtract average frames (END)
IJ.saveAs(
imp_corr_wobg_sub, "Tiff", stackoutpath + separator + imptitle +
"_corrected_womask_substracted.tif")
#pr
IJ.saveAs(
zpimp, "Tiff",
stackoutpath + separator + imptitle + "_corrected_womask_avg.tif")
#commented out: pr
#IJ.saveAs(imp_corr_wobg, "Tiff", stackoutpath+separator+imptitle+"_corrected_womask.tif");
IJ.saveAs(imp_bgmask, "Tiff",
stackoutpath + separator + imptitle + "_bgmask.tif")
print(stackoutpath + separator + imptitle +
"_corrected_womask_substracted.tif")
print(stackoutpath + separator + imptitle + "_corrected_womask_avg.tif")
print(stackoutpath + separator + imptitle + "_bgmask.tif")
imp_corr.changes = False
imp_corr.close()
imp_bgmask.changes = False
imp_bgmask.close()
imp_corr_wobg.changes = False
imp_corr_wobg.close()
#imp_corr_wobg_sub.changes = False
#imp_corr_wobg_sub.close()
zpimp.changes = False
zpimp.close()
#IJ.log(System.getProperty("os.name"))
#IJ.log(fdir)
return imp_corr_wobg_sub
xml = '<?xml version="1.0" encoding="utf-8"?><DyeData>'
profiles=[]
names=[]
for idye_,imp in enumerate(imps):
idye = idye_+1
# Assumes names are FILENAME+" - "+SERIESNAME
names.append(imp.title.split(" - ")[1])
xml+="<Dye%d>"%(idye)
# ZProject
zp = ZProjector(imp)
zp.setMethod(zp.MAX_METHOD)
zp.doProjection()
imp_mi = zp.getProjection()
imp_mip = imp_mi.getProcessor()
# autothreshold using the Triangle method and get the thresholds
imp_mip.setAutoThreshold(AutoThresholder.Method.Triangle,True)
mint = imp_mip.getMinThreshold()
maxt = imp_mip.getMaxThreshold()
# we don't need the MIP anymore
imp_mi.close()
# For each channel, get the mean value above threshold and store it
profile=[]
for ich in range(imp.getNChannels()):
p = imp.getStack().getProcessor(ich+1)
def imageprojector(channels, timelist_unsorted, dirs):
""" Projects .lif timepoints and saves in a common directory,
as well as channel separated directories. """
# Defines in path
path = str(Experiment)
# BF Importer
options = ImporterOptions()
try:
options.setId(path)
except Exception(e):
print str(e)
options.setOpenAllSeries(True)
options.setSplitTimepoints(True)
options.setSplitChannels(True)
imps = BF.openImagePlus(options)
timelist = [x for item in timelist_unsorted for x in repeat(item, channels)]
timelist, imps = zip(*sorted(zip(timelist, imps)))
counter_C0 = -1
counter_C1 = -1
counter_C2 = -1
# Opens all images, splits channels, z-projects and saves to disk
for imp in (imps):
# Projection, Sum Intensity
project = ZProjector()
project.setMethod(ZProjector.SUM_METHOD)
project.setImage(imp)
project.doProjection()
impout = project.getProjection()
projection = impout.getTitle()
try:
# Saves channels to disk,
# add more channels here if desired,
# remember to define new counters.
if "C=0" in projection:
counter_C0 += 1
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections"],
"Scan" + str(counter_C0).zfill(3) + "C0"))
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections_C0"],
"Scan" + str(counter_C0).zfill(3) + "C0"))
elif "C=1" in projection:
counter_C1 += 1
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections"],
"Scan" + str(counter_C1).zfill(3) + "C1"))
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections_C1"],
"Scan" + str(counter_C1).zfill(3) + "C1"))
elif "C=2" in projection:
counter_C2 += 1
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections"],
"Scan" + str(counter_C2).zfill(3) + "C2"))
IJ.saveAs(impout, "TIFF", os.path.join(dirs["Projections_C2"],
"Scan" + str(counter_C2).zfill(3) + "C2"))
except IOException:
print "Directory does not exist"
raise
IJ.log("Images projected and saved to disk")
def CheckMoment(image):
#Perform a maximum projection on the stack
stack = image.getStack()
NSlices=image.getNSlices()
#Take a fixed number of central slices
i=float(NSlices-SimulationCharacteristics['CentralZSlices'])
i1=math.floor(i/2)
i2=math.ceil(i/2)
startSlice=int(1+i1)
stopSlice=int(NSlices-i2)
p=bool(1)
proj = ZProjector()
proj.setMethod(ZProjector.MAX_METHOD)
proj.setImage(image)
proj.setStartSlice(startSlice)
proj.setStopSlice(stopSlice)
proj.doHyperStackProjection(p)
imageMax = proj.getProjection()
imageMax.show()
stack = imageMax.getStack()
n_slices= stack.getSize()
#Get a Segmentation Mask List for every frame in the stack
RoiList=[]
RoiRatioList=[]
for index in range(1, n_slices+1):
ip = stack.getProcessor(index).convertToFloat()
boundRoi=SegmentMask(ip)
mask=boundRoi.getMask()
PixelsMask=mask.getPixels()
r=(-1)*float(sum(PixelsMask))/(ip.height*ip.width)
RoiList.append(boundRoi)
RoiRatioList.append(r)
# print(RoiRatioList)
#Optimize the Roi List. No 1.0 ratios
AllRois1=all(item == 1.0 for item in RoiRatioList)
NFrames=len(RoiRatioList)
if not AllRois1:
for i in range(NFrames):
if RoiRatioList[i]==1.0 or RoiList[i]==None:
#Find the next value that is not 1.0
NearestNot1=-1
prevIndex=i-1
nextIndex=i+1
Token=True #To run through the list alternating positions
while prevIndex>=0 or nextIndex<NFrames:
if prevIndex>0 and Token:
if RoiRatioList[prevIndex]<0.9999:
NearestNot1=prevIndex
break
else:
prevIndex-=1
if nextIndex<NFrames:Token=False
continue
if nextIndex<NFrames:
if RoiRatioList[nextIndex]<0.9999:
NearestNot1=nextIndex
break
else:
nextIndex+=1
if prevIndex>0: Token=True
continue
if NearestNot1!=-1:
#Change RoiList and RoiRationList for the NearestNot1 element
RoiList[i]=RoiList[NearestNot1]
RoiRatioList[i]=RoiRatioList[NearestNot1]
else:
AllRois1=True
print("No avaliable segmentation")
if not AllRois1:
RoiListMin=ModifyRoiList(RoiRatioList,RoiList)
else:
RoiListMin=[ None for i in range(len(RoiList))]
#Find Optimum Scale for the Wavelet
Gamma=endosomeCharacteristics['Gamma_media']
To=endosomeCharacteristics['A_media']
index=1 #Image 1
ipOrig = stack.getProcessor(index).convertToFloat()
boundRoi=RoiListMin[0]
MaxAmp,Ropt=AmplificationCurveMax(ipOrig,To,Gamma,boundRoi)
print("Choosen Scale",Ropt)
Moment3Array=[]
for index in range(1,n_slices+1):
print("time:"+str(index))
ip = stack.getProcessor(index).convertToFloat()
boundRoi=RoiListMin[index-1]
Skew=GetSigmaWavelet(ip,Ropt,boundRoi)
Moment3Array.append(Skew)
return Moment3Array
def maxZprojection(stackimp): zp = ZProjector(stackimp) zp.setMethod(ZProjector.MAX_METHOD) zp.doProjection() zpimp = zp.getProjection() return (zpimp)
def processFile(filename, inDir, outDir, dichroics, mergeList):
if mergeList is None:
merge = False
else:
merge = True
filenameExExt = os.path.splitext(filename)[0]
filepath = inDir + filename
# parse metadata
reader = ImageReader()
omeMeta = MetadataTools.createOMEXMLMetadata()
reader.setMetadataStore(omeMeta)
reader.setId(filepath)
numChannels = reader.getSizeC()
numSlices = reader.getSizeZ()
numFrames = reader.getSizeT()
seriesCount = reader.getSeriesCount()
globalMetadata = reader.getGlobalMetadata()
seriesMetadata = reader.getSeriesMetadata()
objLensName = globalMetadata['- Objective Lens name #1']
areaRotation = float(seriesMetadata['area rotation #1'])
acquisitionValueRotation = float(seriesMetadata['acquisitionValue rotation #1'])
if 'regionInfo rotation #1' in seriesMetadata:
regionInfoRotation = float(seriesMetadata['regionInfo rotation #1'])
else:
regionInfoRotation = float(0)
totalRotation = areaRotation + regionInfoRotation
physSizeX = omeMeta.getPixelsPhysicalSizeX(0)
physSizeY = omeMeta.getPixelsPhysicalSizeY(0)
pxSizeX = physSizeX.value(UNITS.MICROM)
pxSizeY = physSizeY.value(UNITS.MICROM)
# log metadata
IJ.log("\nMETADATA")
#IJ.log("Filename: " + filepath)
IJ.log("Number of series: " + str(seriesCount))
IJ.log("Number of channels: " + str(numChannels))
IJ.log("Number of frames: " + str(numFrames))
IJ.log("Number of slices: " + str(numSlices))
IJ.log("Objective lens: " + objLensName)
IJ.log("FOV rotation: " + str(areaRotation))
IJ.log("ROI rotation: " + str(regionInfoRotation))
IJ.log("Total rotation: " + str(totalRotation))
IJ.log("Pixel size:")
IJ.log("\t\tX = " + str(physSizeX.value()) + " " + physSizeX.unit().getSymbol())
IJ.log("\t\tY = " + str(physSizeY.value()) + " " + physSizeY.unit().getSymbol())
if merge:
tifDir = outDir + "." + str(datetime.now()).replace(" ", "").replace(":", "") + "/"
if not os.path.exists(tifDir):
os.makedirs(tifDir)
IJ.log("\nCreated temporary folder: " + tifDir + "\n")
else:
IJ.log("Unable to create temporary folder!\n")
else:
tifDir = outDir + filenameExExt + "/"
if not os.path.exists(tifDir):
os.makedirs(tifDir)
IJ.log("\nCreated subfolder: " + tifDir + "\n")
else:
IJ.log("\nSubfolder " + tifDir + " already exists.\n")
# correct images
tifFilePaths = []
for i in range(numChannels):
ip = extractChannel(oirFile=filepath, ch=i)
if dichroics[i] == "DM1":
IJ.log("Channel " + str(i+1) + " was imaged using DM1, so no correction required.")
else:
offsets = getOffset(obj=objLensName,dm=dichroicDict[dichroics[i]])
xom = offsets['x']
yom = offsets['y']
if abs(totalRotation) > 0.1:
rotOff = rotateOffset(x=xom, y=yom, angle=-totalRotation)
xom = rotOff['x']
yom = rotOff['y']
xop = int(round(xom/pxSizeX))
yop = int(round(yom/pxSizeY))
IJ.log("Channel " + str(i+1) + " offsets")
IJ.log("\t\tMicrometres")
IJ.log("\t\t\t\tx = " + str(xom))
IJ.log("\t\t\t\ty = " + str(yom))
IJ.log("\t\tPixels")
IJ.log("\t\t\t\tx = " + str(xop))
IJ.log("\t\t\t\ty = " + str(yop))
IJ.run(ip, "Translate...", "x=" + str(-xop) + " y=" + str(-yop) + " interpolation=None stack")
tifFilePath = tifDir + filenameExExt + "_ch_"+str(i+1)+".tif"
tifFilePaths.append(tifFilePath)
if os.path.exists(tifFilePath):
IJ.log("\nOutput file exists: " + tifFilePath)
IJ.log("Rerun plugin choosing a different output folder")
IJ.log("or delete file and then rerun plugin.")
IJ.log("Image processing terminated!\n")
return
FileSaver(ip).saveAsTiff(tifFilePath)
if merge:
max_list = []
for i in range(len(mergeList)):
if mergeList[i] != None:
mergeList[i] = readSingleChannelImg(tifFilePaths[mergeList[i]])
channel = mergeList[i]#https://python.hotexamples.com/examples/ij.plugin/RGBStackMerge/mergeChannels/python-rgbstackmerge-mergechannels-method-examples.html
projector = ZProjector(channel)
projector.setMethod(ZProjector.MAX_METHOD)
projector.doProjection()
max_list.append(projector.getProjection())
merged = RGBStackMerge.mergeChannels(mergeList, False)
merged_max = RGBStackMerge.mergeChannels(max_list, False)
mergedChannelFilepath = outDir + filenameExExt + ".tif"
maxMergedChannelFilepath = outDir + filenameExExt + "_max.tif"
if os.path.exists(mergedChannelFilepath):
IJ.log("\nOutput file exists: " + mergedChannelFilepath)
IJ.log("Rerun plugin choosing a different output folder")
IJ.log("or delete file and then rerun plugin.")
IJ.log("Image processing terminated!\n")
FileSaver(merged).saveAsTiff(mergedChannelFilepath)
FileSaver(merged_max).saveAsTiff(maxMergedChannelFilepath)
for tf in tifFilePaths:
os.remove(tf)
os.rmdir(tifDir)
IJ.log("\nFinished processing file:\n" + filepath + "\n")
if merge:
IJ.log("Image file with channels aligned:\n" + outDir + filenameExExt + ".tif\n")
else:
IJ.log("Aligned images (one tiff file for each channel) can be found in:\n" + tifDir + "\n")
def ColMigBud():
def setupDialog(imp):
gd = GenericDialog("Collective migration buddy options")
gd.addMessage("Collective migration buddy 2.0, you are analyzing: " +
imp.getTitle())
calibration = imp.getCalibration()
if (calibration.frameInterval > 0):
default_interval = calibration.frameInterval
default_timeunit = calibration.getTimeUnit()
else:
default_interval = 8
default_timeunit = "min"
gd.addNumericField("Frame interval:", default_interval,
2) # show 2 decimals
gd.addCheckbox("Do you want to use a gliding window?", True)
gd.addCheckbox(
"Project hyperStack? (defaluts to projecting current channel only)",
False)
gd.addStringField("time unit", default_timeunit, 3)
gd.addSlider("Start compacting at frame:", 1, imp.getNFrames(), 1)
gd.addSlider("Stop compacting at frame:", 1, imp.getNFrames(),
imp.getNFrames())
gd.addNumericField("Number of frames to project in to one:", 3,
0) # show 0 decimals
gd.addChoice('Method to use for frame projection:', methods_as_strings,
methods_as_strings[5])
gd.showDialog()
if gd.wasCanceled():
IJ.log("User canceled dialog!")
return
return gd
#Start by getting the active image window and get the current active channel and other stats
imp = WindowManager.getCurrentImage()
cal = imp.getCalibration()
nSlices = 1 #TODO fix this in case you want to do Z-stacks
title = imp.getTitle()
current_channel = imp.getChannel()
#zp = ZProjector(imp)
#Make a dict containg method_name:const_fieled_value pairs for the projection methods
methods_as_strings = [
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices',
'Standard Deviation', 'Median'
]
methods_as_const = [
ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD,
ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD
]
medthod_dict = dict(zip(methods_as_strings, methods_as_const))
# Run the setupDialog, read out and store the options
gd = setupDialog(imp)
frame_interval = gd.getNextNumber()
time_unit = gd.getNextString()
glidingFlag = gd.getNextBoolean()
hyperstackFlag = gd.getNextBoolean()
#Set the frame interval and unit, and store it in the ImagePlus calibration
cal.frameInterval = frame_interval
cal.setTimeUnit(time_unit)
imp.setCalibration(cal)
start_frame = int(gd.getNextNumber())
stop_frame = int(gd.getNextNumber())
#If a subset of the image is to be projected, these lines of code handle that
if (start_frame > stop_frame):
IJ.showMessage("Start frame > Stop frame, can't go backwards in time!")
raise RuntimeException("Start frame > Stop frame!")
if ((start_frame != 1) or (stop_frame != imp.getNFrames())):
imp = Duplicator().run(imp, 1, nChannels, 1, nSlices, start_frame,
stop_frame)
no_frames_per_integral = int(gd.getNextNumber())
#the doHyperstackProjection method can't project past the end of the stack
if hyperstackFlag:
total_no_frames_to_project = imp.getNFrames() - no_frames_per_integral
#the doProjection method can project past the end, it just adds black frames at the end
#When not projecting hyperstacks, just copy the current active channel from the active image
else:
total_no_frames_to_project = imp.getNFrames()
imp = Duplicator().run(imp, current_channel, current_channel, 1,
nSlices, start_frame, stop_frame)
#The Z-Projection magic happens here through a ZProjector object
zp = ZProjector(imp)
projection_method = gd.getNextChoice()
chosen_method = medthod_dict[projection_method]
zp.setMethod(chosen_method)
outstack = imp.createEmptyStack()
if glidingFlag:
frames_to_advance_per_step = 1
else:
frames_to_advance_per_step = no_frames_per_integral
for frame in range(1, total_no_frames_to_project,
frames_to_advance_per_step):
zp.setStartSlice(frame)
zp.setStopSlice(frame + no_frames_per_integral)
if hyperstackFlag:
zp.doHyperStackProjection(False)
projected_stack = zp.getProjection().getStack()
for channel in range(projected_stack.getSize()):
outstack.addSlice(projected_stack.getProcessor(channel + 1))
else:
zp.doProjection()
outstack.addSlice(zp.getProjection().getProcessor())
#Create an image processor from the newly created Z-projection stack
nChannels = imp.getNChannels()
nFrames = outstack.getSize() / nChannels
imp2 = ImagePlus(
title + '_' + projection_method + '_' + str(no_frames_per_integral) +
'_frames', outstack)
imp2 = HyperStackConverter.toHyperStack(imp2, nChannels, nSlices, nFrames)
imp2.show()
Startmenu()
def maxZprojection(self, inputimg):
zp = ZProjector(inputimg)
zp.setMethod(ZProjector.MAX_METHOD)
zp.doProjection()
Zproj = zp.getProjection()
return Zproj
def BackgroundFilter():
def setupDialog(imp):
gd = GenericDialog("Toolbox options")
gd.addMessage("Imagej toolbox, you are analyzing: " + imp.getTitle())
calibration = imp.getCalibration()
if (calibration.frameInterval > 0):
default_interval = calibration.frameInterval
default_timeunit = calibration.getTimeUnit()
else:
default_interval = 10
default_timeunit = "sec"
gd.addNumericField("Frame interval:", default_interval,
2) # show 2 decimals
# gd.addCheckbox("Do you want to use a gliding window?", True)
# gd.addCheckbox("Project hyperStack? (defaluts to projecting current channel only)", False)
# gd.addStringField("time unit",default_timeunit, 3)
# gd.addSlider("Start compacting at frame:", 1, imp.getNFrames(), 1)
# gd.addSlider("Stop compacting at frame:", 1, imp.getNFrames(), imp.getNFrames())
# gd.addNumericField("Number of frames to project in to one:", 3, 0) # show 0 decimals
gd.addChoice('Method to use for stack background filtering:',
methods_as_strings, methods_as_strings[5])
gd.showDialog()
if gd.wasCanceled():
IJ.log("User canceled dialog!")
return
return gd
#Start by getting the active image window and get the current active channel and other stats
imp = WindowManager.getCurrentImage()
cal = imp.getCalibration()
nSlices = 1 #TODO fix this in case you want to do Z-stacks
title = imp.getTitle()
current_channel = imp.getChannel()
#zp = ZProjector(imp)
#Make a dict containg method_name:const_fieled_value pairs for the projection methods
methods_as_strings = [
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices',
'Standard Deviation', 'Median'
]
methods_as_const = [
ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD,
ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD
]
method_dict = dict(zip(methods_as_strings, methods_as_const))
gd = setupDialog(imp)
#The Z-Projection magic happens here through a ZProjector object
zp = ZProjector(imp)
projection_method = gd.getNextChoice()
chosen_method = method_dict[projection_method]
zp.setMethod(chosen_method)
outstack = imp.createEmptyStack()
zp.doProjection()
outstack.addSlice(zp.getProjection().getProcessor())
imp2 = ImagePlus(title + '_' + projection_method, outstack)
# imp2.show()
imp3 = ImageCalculator().run("Subtract create 32-bit stack", imp, imp2)
imp3.show()
Startmenu()
def glidingprojection(imp,
startframe=1,
stopframe=None,
glidingFlag=True,
no_frames_per_integral=3,
projectionmethod="Median"):
"""This function subtracts the gliding projection of several frames from the
input stack. Thus, everything which moves too fast is filtered away.
Args:
imp (ImagePlus): Input image as ImagePlus object.
startframe (int, optional): Choose a start frame. Defaults to 1.
stopframe (int, optional): Choose an end frame. Defaults to None.
glidingFlag (bool, optional): Should a gliding frame by frame projection be used? Defaults to True.
no_frames_per_integral (int, optional): Number of frames to project each integral. Defaults to 3.
projectionmethod (str, optional): Choose the projection method. Options are
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices', 'Standard Deviation', 'Median'. Defaults to "Median".
Raises:
RuntimeException: Start frame > stop frame.
Returns:
ImagePlus: The output stack.
"""
# Store some image properties.
cal = imp.getCalibration()
width, height, nChannels, nSlices, nFrames = imp.getDimensions()
title = imp.getTitle()
# Some simple sanity checks for input parameters.
if stopframe == None: stopframe = nFrames
if (startframe > stopframe):
IJ.showMessage("Start frame > Stop frame, can't go backwards in time!")
raise RuntimeException("Start frame > Stop frame!")
# If a subset of the image is to be projected, these lines of code handle that.
if ((startframe != 1) or (stopframe != nFrames)):
imp = Duplicator().run(imp, 1, nChannels, 1, nSlices, startframe,
stopframe)
# Define the number of frames to advance per step based on boolean input parameter glidingFlag.
if glidingFlag: frames_to_advance_per_step = 1
else: frames_to_advance_per_step = no_frames_per_integral
# Make a dict containg method_name:const_fieled_value pairs for the projection methods
methods_as_strings = [
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices',
'Standard Deviation', 'Median'
]
methods_as_const = [
ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD,
ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD
]
method_dict = dict(zip(methods_as_strings, methods_as_const))
# Initialize a ZProjector object and an empty stack to collect projections.
zp = ZProjector(imp)
zp.setMethod(method_dict[projectionmethod])
outstack = imp.createEmptyStack()
# Loop through all the frames in the image, and project that frame with the other frames in the integral.
for frame in range(1, nFrames + 1, frames_to_advance_per_step):
zp.setStartSlice(frame)
zp.setStopSlice(frame + no_frames_per_integral)
zp.doProjection()
outstack.addSlice(zp.getProjection().getProcessor())
# Create an image processor from the newly created Z-projection stack
# nFrames = outstack.getSize()/nChannels
impout = ImagePlus(
title + '_' + projectionmethod + '_' + str(no_frames_per_integral) +
'_frames', outstack)
impout = HyperStackConverter.toHyperStack(impout, nChannels, nSlices,
nFrames)
impout.setCalibration(cal)
return impout
# improved with default values or min/max values.
emissionWL = [0]
try:
for x in xrange(1,nbrChannels+1,1):
WL = int(imp.getNumericProperty("Information|Image|Channel|EmissionWavelength #" + str(x).zfill(2)))
emissionWL.append(WL)
except ValueError:
raise Exception("Couldn't find the EmissionWavelength info in the metadata")
# Make the maximum intensity projection
# and show it for user input.
project = ZProjector()
project.setMethod(ZProjector.MAX_METHOD)
project.setImage(imp)
project.doProjection()
projection = project.getProjection()
projection.show()
# Wait for the user to draw a line
# and check if line ROI.
IJ.setTool("line");
myWait = WaitForUserDialog ("waitForUser", "Make a line with the region of interest and press OK")
myWait.show()
roi = projection.getRoi()
# Check if roi is a line
# otherwise exit
if not (roi.isLine()):
raise Exception("Line selection required")
if options is not None:
runCamR, runCamL, CamRScale, CamLScale, BackgroundWindowStart, BackgroundWindowEnd, GridSize, TranslateR, TranslateL = options # unpack each parameter
print runCamR, runCamL, CamRScale, CamLScale, BackgroundWindowStart, BackgroundWindowEnd, GridSize, TranslateR, TranslateL
if runCamR:
impR1 = FolderOpener.open(folder, "file=CamR sort")
UID = impR1.title
filepath = folder.strip(UID + '/')
projectR = ZProjector(impR1)
projectR.setMethod(ZProjector.AVG_METHOD)
projectR.setImage(impR1)
projectR.setStartSlice(int(BackgroundWindowStart))
projectR.setStopSlice(int(BackgroundWindowEnd))
projectR.doProjection()
projectionR = projectR.getProjection()
impR2 = ImageCalculator().run("Subtract create stack", impR1, projectionR)
IJ.run(impR2, "Enhance Contrast", "saturated=0.4 process_all")
IJ.run(impR2, "Set Scale...",
"distance=" + str(CamRScale) + " known=1 unit=cm")
IJ.run(impR2, "Grid...",
"grid=Lines area=" + str(GridSize) + " color=Cyan")
IJ.run(impR2, "Translate...",
"x=" + str(TranslateR) + " y=0 interpolation=None stack")
savepath = path.join(folder, UID + "_CR.tif")
print savepath
fs = FileSaver(impR2)
if fs.saveAsTiffStack(savepath):
print("Files saved succesfully")
#print "index=" + str(index) + "start=" + str(currStart) + " stop=" + str(currStop)
if (currStart<1):
currStart = 1
if (currStop>nSlices):
currStop = nSlices
zp = ZProjector(imp)
zp.setMethod(ZProjector.MAX_METHOD)
zp.setStartSlice(currStart)
zp.setStopSlice(currStop)
#zp.doHyperStackProjection(True)
zp.doProjection()
tmpProject = zp.getProjection()
zImageProcessor = tmpProject.getProcessor()
if (tmpProject==None):
print "\tNULL ZProject"
else:
newStack.addSlice(origStack.getSliceLabel(index), zImageProcessor)
currStart = index - userSlices
currStop = index + userSlices
#how the results as an image
imp2 = ImagePlus("Sliding Z Project: " + imp.title + " " + "[" + str(userSlices) + "]", newStack)
imp2.setCalibration(imp.getCalibration().copy())
imp2.show()