def scaleandfilter(infile,outfile,scalex,scaley):
print ("infile is: "+infile)
imp = Opener().openImage(infile)
print imp
print "scalex = %f; scaley = %f" % (scalex,scaley)
# Rescale
ip = imp.getProcessor()
ip.setInterpolate(True)
sp = StackProcessor(imp.getStack(),ip);
sp2=sp.resize(int(round(ip.width * scalex)), int(round(ip.height *scaley)));
imp.setStack(imp.getTitle(),sp2);
cal = imp.getCalibration()
cal.pixelWidth /= scalex
cal.pixelHeight /= scaley
IJ.run(imp, "8-bit","")
intif=infile+".tif"
outtif=infile+"-filtered.tif"
print("saving input file as "+intif)
f=FileSaver(imp)
f.saveAsTiffStack(intif)
imp.close()
# anisotropic filtering
anisopts="-scanrange:10 -tau:2 -nsteps:2 -lambda:0.1 -ipflag:0 -anicoeff1:1 -anicoeff2:0 -anicoeff3:0"
anisopts=anisopts+" -dx:%f -dy:%f -dz:%f" % (cal.pixelWidth,cal.pixelHeight,cal.pixelDepth)
if sys.version_info > (2, 4):
#for testing
# subprocess.check_call(["cp",intif,outtif])
subprocess.check_call(["anisofilter"]+anisopts.split(' ')+[intif,outtif])
else:
os.system(" ".join(["anisofilter"]+anisopts.split(' ')+[intif,outtif]))
# Hessian (tubeness)
print("Opening output tif: "+outtif)
imp = Opener().openImage(outtif)
imp.setCalibration(cal)
print("Running tubeness on tif: "+outtif)
IJ.run(imp,"Tubeness", "sigma=1")
IJ.run(imp, "8-bit","")
# Save to PIC
print("Saving as PIC: "+outfile)
# IJ.saveAs("tiff","outtif")
IJ.run(imp,"Biorad ...", "biorad="+outfile)
def scaleandfilter(infile,outfile,scalex,scaley,scalez,anisofilter,runtube):
print ("infile is: "+infile)
imp = Opener().openImage(infile)
print imp
print "scalex = %f; scaley = %f ; scalez = %f" % (scalex,scaley,scalez)
# Rescale
cal = imp.getCalibration()
iml = ImgLib.wrap(imp)
scaledimg = Scale3D(iml, scalex, scaley, scalez)
imp2=ImgLib.wrap(scaledimg)
# find range of pixel values for scaled image
from mpicbg.imglib.algorithm.math import ComputeMinMax
# (for imglib2 will be: net.imglib2.algorithm.stats)
minmax=ComputeMinMax(scaledimg)
minmax.process()
(min,max)=(minmax.getMin().get(),minmax.getMax().get())
# Make a copy of the stack (converting to 8 bit as we go)
stack = ImageStack(imp2.width, imp2.height)
print "min = %e, max =%e" % (min,max)
for i in xrange(1, imp2.getNSlices()+1):
imp2.setSliceWithoutUpdate(i)
ip=imp2.getProcessor()
# set range
ip.setMinAndMax(min,max)
stack.addSlice(str(i), ip.convertToByte(True))
# save copy of calibration info
cal=imp.getCalibration()
# close original image
imp.close()
# make an image plus with the copy
scaled = ImagePlus(imp2.title, stack)
# Deal with calibration info which didn't seem to come along for the ride
cal.pixelWidth/=scalex
cal.pixelHeight/=scaley
cal.pixelDepth/=scalez
scaled.setCalibration(cal)
print "dx = %f; dy=%f; dz=%f" % (cal.pixelWidth,cal.pixelHeight,cal.pixelDepth)
intif=infile+".tif"
outtif=infile+"-filtered.tif"
if anisofilter.upper() != 'FALSE':
print("saving input file as "+intif)
f=FileSaver(scaled)
f.saveAsTiffStack(intif)
scaled.close()
# anisotropic filtering
anisopts="-scanrange:10 -tau:2 -nsteps:2 -lambda:0.1 -ipflag:0 -anicoeff1:1 -anicoeff2:0 -anicoeff3:0"
anisopts=anisopts+" -dx:%f -dy:%f -dz:%f" % (cal.pixelWidth,cal.pixelHeight,cal.pixelDepth)
if sys.version_info > (2, 4):
#for testing
# subprocess.check_call(["cp",intif,outtif])
subprocess.check_call([anisofilter]+anisopts.split(' ')+[intif,outtif])
else:
os.system(" ".join([anisofilter]+anisopts.split(' ')+[intif,outtif]))
# Open anisofilter output back into Fiji
print("Opening output tif: "+outtif)
scaled = Opener().openImage(outtif)
scaled.setCalibration(cal)
# Hessian (tubeness)
print("Running tubeness")
if(runtube):
tp=TubenessProcessor(1.0,False)
result = tp.generateImage(scaled)
IJ.run(result, "8-bit","")
else:
result=scaled
# Save out file
fileName, fileExtension = os.path.splitext(outfile)
print("Saving as "+fileExtension+": "+outfile)
if fileExtension.lower()=='.nrrd':
nw=Nrrd_Writer()
nw.setNrrdEncoding("gzip")
nw.save(result,outfile)
else:
# Save to PIC
IJ.run(result,"Biorad ...", "biorad=["+outfile+"]")
scaled.close()
result.close()
class bImp:
def __init__(self, filepath):
"""
Load an image or stack from filepath.
Args:
filepath (str): Full path to an image file. Can be .tif, .lsm, .czi, etc
"""
if not os.path.isfile(filepath):
bPrintLog('ERROR: bImp() did not find file: ' + filepath,0)
return 0
self.filepath = filepath
folderpath, filename = os.path.split(filepath)
self.filename = filename
self.enclosingPath = folderpath
self.enclosingfolder = os.path.split(folderpath)[1]
self.dateStr = ''
self.timeStr = ''
self.imp = None
tmpBaseName, extension = os.path.splitext(filename)
isZeiss = extension in ['.czi', '.lsm']
self.islsm = extension == '.lsm'
self.isczi = extension == '.czi'
istif = extension == '.tif'
if istif:
# scanimage3 comes in with dimensions: [512, 512, 1, 52, 1]) = [width, height, numChannels, numSlices, numFrames]
self.imp = Opener().openImage(filepath)
self.imp.show()
elif isZeiss:
#open lsm using LOCI Bio-Formats
options = ImporterOptions()
#options.setColorMode(ImporterOptions.COLOR_MODE_GRAYSCALE)
options.setId(filepath)
imps = BF.openImagePlus(options)
for imp in imps:
self.imp = imp #WindowManager.getImage(self.windowname)
imp.show()
if not self.imp:
bPrintLog('ERROR: bImp() was not able to open file: '+ filepath,0)
self.windowname = filename
#self.imp = WindowManager.getImage(self.windowname)
# numChannels is not correct for scanimage, corrected in readTiffHeader()
(width, height, numChannels, numSlices, numFrames) = self.imp.getDimensions()
self.width = width # pixelsPerLine
self.height = height # linesPerFrame
self.numChannels = numChannels
self.numSlices = numSlices
self.numFrames = numFrames
self.infoStr = self.imp.getProperty("Info") #get all tags
self.voxelx = 1
self.voxely = 1
self.voxelz = 1
#self.numChannels = 1
#self.bitsPerPixel = 8
self.zoom = 1
self.motorx = None
self.motory = None
self.motorz = None
self.scanImageVersion = ''
self.msPerLine = None
self.dwellTime = None
# read file headers (date, time, voxel size)
if isZeiss:
self.readZeissHeader(self.infoStr)
elif istif:
self.readTiffHeader(self.infoStr)
self.updateInfoStr()
self.channelWindows = []
self.channelImp = []
if self.numChannels == 1:
self.channelWindows.append(self.windowname)
self.channelImp.append(self.imp)
else:
self.deinterleave()
def updateInfoStr(self):
# Fill in infoStr with Map Manager tags
self.infoStr += 'Folder2MapManager=' + versionStr + '\n'
self.infoStr += 'b_date=' + self.dateStr + '\n'
self.infoStr += 'b_time=' + self.timeStr + '\n'
# yevgeniya 20180314
#if (self.numChannels > 3):
# self.numChannels = 3
self.infoStr += 'b_numChannels=' + str(self.numChannels) + '\n'
self.infoStr += 'b_pixelsPerline=' + str(self.width) + '\n'
self.infoStr += 'b_linesPerFrame=' + str(self.height) + '\n'
self.infoStr += 'b_numSlices=' + str(self.numSlices) + '\n'
self.infoStr += 'b_voxelX=' + str(self.voxelx) + '\n'
self.infoStr += 'b_voxelY=' + str(self.voxely) + '\n'
self.infoStr += 'b_voxelZ=' + str(self.voxelz) + '\n'
#self.infoStr += 'b_bitsPerPixel=' + str(self.bitsPerPixel) + '\n'
self.infoStr += 'b_zoom=' + str(self.zoom) + '\n'
self.infoStr += 'b_motorx=' + str(self.motorx) + '\n'
self.infoStr += 'b_motory=' + str(self.motory) + '\n'
self.infoStr += 'b_motorz=' + str(self.motorz) + '\n'
self.infoStr += 'b_msPerLine=' + str(self.msPerLine) + '\n'
self.infoStr += 'b_scanImageVersion=' + self.scanImageVersion + '\n'
def readTiffHeader(self, infoStr):
"""
Read ScanImage 3/4 .tif headers
"""
logLevel = 3
# splitting on '\r' for scanimage 3.x works
# splitting on '\n' for scanimage 4.x works
#we need to search whole infoStr to figure out scanimage 3 or 4.
# we can't split info string because si3 uses \r and si4 uses \n
infoStrDelim = '\n'
if infoStr.find('scanimage.SI4') != -1:
infoStrDelim = '\n'
bPrintLog('Assuming SI4 infoStr to be delimited with backslash n', logLevel)
elif infoStr.find('state.software.version') != -1:
infoStrDelim = '\r'
bPrintLog('Assuming SI3 infoStr to be delimited with backslash r', logLevel)
else:
bPrintLog('Splitting infoStr using backslah n', logLevel)
# if we don't find zoom then voxel is an error (see end of function)
foundZoom = False
#for line in infoStr.split('\n'):
for line in infoStr.split(infoStrDelim):
#
# ScanImage 4.x
#
# scanimage.SI4.versionMajor = 4.2
if line.find('scanimage.SI4.versionMajor') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.scanImageVersion = rhs
# scanimage.SI4.motorPosition = [-33936.5 -106316 -55308.5]
if line.find('scanimage.SI4.motorPosition') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
rhs = rhs.replace('[','')
rhs = rhs.replace(']','')
floats = [float(x) for x in rhs.split()]
self.motorx = floats[0]
self.motory = floats[1]
self.motorz = floats[2]
# scanimage.SI4.channelsSave = [1;2]
if line.find('scanimage.SI4.channelsSave') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
rhs = rhs.replace('[','')
rhs = rhs.replace(']','')
channels = [int(x) for x in rhs.split(';')]
bPrintLog('reading scanimage.SI4.channelsSave inferred channels:' + str(channels), logLevel)
self.numChannels = len(channels)
# scanimage.SI4.scanZoomFactor = 5.9
if line.find('scanimage.SI4.scanZoomFactor') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.zoom = float(rhs)
foundZoom = True
#self.voxelx = magic_scan_image_scale / self.zoom
#self.voxely = magic_scan_image_scale / self.zoom
# scanimage.SI4.triggerClockTimeFirst = '18-05-2015 11:58:43.788'
if line.find('scanimage.SI4.triggerClockTimeFirst') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
rhs = rhs.replace("'","") # remove enclosing ' and '
if rhs.startswith(' '): # if date string starts with space, remove it
rhs = rhs[1:-1]
datetime = rhs.split(' ')
# 20170811, there is an extra f*****g space before datestr on the rhs
# convert mm/dd/yyyy to yyyymmdd
#print 'rhs:' + "'" + rhs + "'"
#print 'datetime:', datetime
datestr = bFixDate(datetime[0], logLevel)
self.dateStr = datestr
self.timeStr = datetime[1]
#
# ScanImage 3.x
#
# state.software.version = 3.8
if line.find('state.software.version') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.scanImageVersion = rhs
# state.acq.numberOfChannelsAcquire = 2
if line.find('state.acq.numberOfChannelsAcquire') != -1:
#print '\rDEBUG 12345'
bPrintLog(line, logLevel)
#print '\rDEBUG 12345'
rhs = line.split('=')[1]
self.numChannels = int(rhs)
# state.acq.zoomFactor = 2.5
if line.find('state.acq.zoomFactor') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.zoom = float(rhs)
foundZoom = True
# set (voxelx, voxely)
#self.voxelx = magic_scan_image_scale / self.zoom
#self.voxely = magic_scan_image_scale / self.zoom
# state.acq.msPerLine = 2.32
if line.find('state.acq.msPerLine') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.msPerLine = float(rhs)
# state.acq.pixelTime = 3.2e-06
if line.find('state.acq.pixelTime') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.dwellTime = float(rhs)
# state.motor.absXPosition = -9894.4
if line.find('state.motor.absXPosition') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.motorx = float(rhs)
# state.motor.absYPosition = -18423.4
if line.find('state.motor.absYPosition') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.motory = float(rhs)
# state.motor.absZPosition = -23615.04
if line.find('state.motor.absZPosition') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.motorz = float(rhs)
# state.acq.zStepSize = 2
if line.find('state.acq.zStepSize') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
self.voxelz = float(rhs)
# state.internal.triggerTimeString = '10/2/2014 12:29:22.796'
if line.find('state.internal.triggerTimeString') != -1:
bPrintLog(line, logLevel)
rhs = line.split('=')[1]
rhs = rhs.replace("'","")
if rhs.startswith(' '): # if date string starts with space, remove it
rhs = rhs[1:-1]
datetime = rhs.split(' ')
# 20170811, there is an extra f*****g space before datestr on the rhs
# convert mm/dd/yyyy to yyyymmdd
#print 'rhs:' + "'" + rhs + "'"
#print 'datetime:', datetime
self.dateStr = bFixDate(datetime[0], logLevel)
self.timeStr = bFixTime(datetime[1], logLevel)
# state.acq.acqDelay = 0.000122
# state.acq.bidirectionalScan = 0
# state.acq.fillFraction = 0.706206896551724
# state.acq.frameRate = 0.841864224137931
# huganir lab keeps this off, image pixel intensities are 2^11 * samplesperpixel (e.g. binFactor?)
# state.acq.binFactor = 16
# state.internal.averageSamples = 1
# the real image bit depth is usually inputBitDepth-1 (1 bit is not used?)
# state.acq.inputBitDepth = 12
if foundZoom:
self.voxelx = magic_scan_image_scale / self.zoom * (1024 / self.width)
self.voxely = magic_scan_image_scale / self.zoom * (1024 / self.height)
else:
bPrintLog('ERROR: Did not find zoom in SI header, voxel x/y will be wrong', logLevel)
def readZeissHeader(self, infoStr):
# This is incredibly difficult to get working as (date, time, voxels) are in different obscure places in lsm and czi
# Furthermore, just trying to read the raw ome xls is futile
#
# parsing ome xml as a string and searching it with regular expression(re) does not work
# it is beyond the scope of my work to figure this out
# the fact that it does not work and there is little documentaiton is a pretty big waste of time
#
# get and parse xml to find date/time
#fi = self.imp.getOriginalFileInfo(); # returns a FileInfo object
#omexml = fi.description #omexml is a string
#omexml = omexml.encode('utf-8')
#omexml = omexml.replaceAll("[^\\x20-\\x7e]", "") # see: https://stackoverflow.com/questions/2599919/java-parsing-xml-document-gives-content-not-allowed-in-prolog-error
# (1) try and search the ome xml like a string, this gives errors
#docsPattern = '<AcquisitionDate>.*</AcquisitionDate>'
#searchresult = re.search(docsPattern, omexml)
#print 'searchresult:', searchresult.group(0)
# 2) treat the ome xml like any other xml (because it's xml, right?)
# well this raises errors too
#omexml has <AcquisitionDate>2016-08-17T15:21:50</AcquisitionDate>
#import xml.etree.ElementTree
#e = xml.etree.ElementTree.fromstring(omexml).getroot() #print omexml
#for atype in e.findall('AcquisitionDate'):
# print 'AcquisitionDate:', atype #.get('foobar')
#
#
if self.islsm:
# lsm have date hidden in omeMeta.getImageAcquisitionDate(0)
# this is copied from code at: https://gist.github.com/ctrueden/6282856
reader = ImageReader()
omeMeta = MetadataTools.createOMEXMLMetadata() #omeMeta.getImageAcquisitionDate(0)
reader.setMetadataStore(omeMeta)
reader.setId(self.filepath)
#seriesCount = reader.getSeriesCount()
dateTimeStr = omeMeta.getImageAcquisitionDate(0) #2016-08-17T16:36:26
reader.close()
if dateTimeStr:
self.dateStr, self.timeStr = dateTimeStr.toString().split('T')
self.dateStr = bFixDate(self.dateStr)
self.timeStr = bFixTime(self.timeStr)
#bPrintLog('LSM date/time is: ' + self.dateStr + ' ' + self.timeStr, 3)
else:
bPrintLog('WARNING: did not get Zeiss date/time string')
# lsm have voxels in infoStr
for line in infoStr.split('\n'):
#print line
if line.find('VoxelSizeX') != -1:
self.voxelx = float(line.split('=')[1])
if line.find('VoxelSizeY') != -1:
self.voxely = float(line.split('=')[1])
if line.find('VoxelSizeZ') != -1:
self.voxelz = float(line.split('=')[1])
if line.find('SizeC') != -1:
self.numChannels = int(line.split('=')[1])
#if line.find('BitsPerPixel') and not line.startswith('Experiment') != -1: # 20170811, startswith is for czi
# self.bitsPerPixel = int(line.split('=')[1])
if line.find('RecordingZoomX#1') != -1:
self.zoom = int(line.split('=')[1])
if self.isczi:
# czi has date/time in infoStr (lsm does not)
for line in infoStr.split('\n'):
if line.find('CreationDate #1') != -1: # w.t.f. is #1 referring to?
lhs, rhs = line.split('=')
rhs = rhs.replace(' ', ' ')
if rhs.startswith(' '):
rhs = rhs[1:-1]
#print "lhs: '" + lhs + "'" + "rhs: '" + rhs + "'"
if rhs.find('T') != -1:
self.dateStr, self.timeStr = rhs.split('T')
else:
self.dateStr, self.timeStr = rhs.split(' ')
self.dateStr = bFixDate(self.dateStr)
self.timeStr = bFixTime(self.timeStr)
#bPrintLog('CZI date/time is: ' + self.dateStr + ' ' + self.timeStr, 3)
# .czi
# <Pixels BigEndian="false" DimensionOrder="XYCZT" ID="Pixels:0" Interleaved="false" PhysicalSizeX="0.20756645602494875" PhysicalSizeXUnit="µm" PhysicalSizeY="0.20756645602494875" PhysicalSizeYUnit="µm" PhysicalSizeZ="0.75" PhysicalSizeZUnit="µm" SignificantBits="8" SizeC="1" SizeT="1" SizeX="1024" SizeY="1024" SizeZ="50" Type="uint8">
# czi have voxel in calibration
self.voxelx = self.imp.getCalibration().pixelWidth;
self.voxely = self.imp.getCalibration().pixelHeight;
self.voxelz = self.imp.getCalibration().pixelDepth;
#bPrintLog('readZeissHeader() read czi scale as: ' + str(self.voxelx) + ' ' + str(self.voxely) + ' ' + str(self.voxelz), 3)
# CLEARING self.infoStr for CZI ... it was WAY to big to parse in Map Manager
self.infoStr = ''
def printParams(self, loglevel=3): # careful, thefunction print() is already taken?
bPrintLog('file:' + self.filepath, loglevel)
bPrintLog("date:'" + self.dateStr + "' time:'" + self.timeStr + "'", loglevel)
bPrintLog('channels:' + str(self.numChannels), loglevel)
bPrintLog('zoom:' + str(self.zoom), loglevel)
bPrintLog('pixels:' + str(self.width) + ',' + str(self.height)+ ',' + str(self.numSlices), loglevel)
bPrintLog('voxels:' + str(self.voxelx) + ',' + str(self.voxely)+ ',' + str(self.voxelz), loglevel)
def deinterleave(self):
if self.numChannels == 1:
bPrintLog('Warning: deinterleave() did not deinterleave with num channels 1', 0)
return -1
#IJ.run('Deinterleave', 'how=' + str(self.numChannels) +' keep') #makes ' #1' and ' #2', with ' #2' frontmost
cmdStr = 'how=' + str(self.numChannels) + ' keep'
IJ.run('Deinterleave', cmdStr) #makes ' #1' and ' #2', with ' #2' frontmost
for i in range(self.numChannels):
currenChannel = i + 1
currentWindowName = self.windowname + ' #' + str(currenChannel)
self.channelWindows.append(currentWindowName)
currentImp = WindowManager.getImage(currentWindowName)
if currentImp:
self.channelImp.append(currentImp)
else:
bPrintLog('ERROR: deinterleave() did not find window names:' + currentWindowName, 0)
def exportTifStack(self, destFolder=''):
channelNumber = 1
for imp in self.channelImp:
if not destFolder:
destFolder = os.path.join(self.enclosingPath, self.enclosingfolder + '_tif')
if not os.path.isdir(destFolder):
os.makedirs(destFolder)
if not imp:
bPrintLog("ERROR: exportTifStack() did not find an imp at channel number '" + str(channelNumber) + "'", 0)
return -1
self.updateInfoStr()
imp.setProperty("Info", self.infoStr);
saveFile = os.path.splitext(self.filename)[0] + '_ch' + str(channelNumber) + '.tif'
savePath = os.path.join(destFolder, saveFile)
# save
fs = FileSaver(imp)
bPrintLog('saveTifStack():' + savePath, 3)
if imp.getNSlices()>1:
fs.saveAsTiffStack(savePath)
else:
fs.saveAsTiff(savePath)
channelNumber += 1
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 closeAll(self):
self.imp.close()
for imp in self.channelImp:
imp.close()
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)
if regEx.match(fileName): # if matches RE, add to list
moFileList.append(regEx.match(fileName))
print "Will process files ", moFileList
if moFileList == []:
IJ.showMessage("Input Exception", "Directory does not contain any preprocessed images.")
raise IOError("Input Exception: Directory does not contain any preprocessed images.")
if not pth.exists(saveFolder): # check if directory for analysis-files is present
makedirs(saveFolder)
for image in moFileList: # get rid of 0!
print "starting with cell " + image.group() + " " + "("+ str(moFileList.index(image)) + "/" + str(len(moFileList)) + ")"
imp = Opener().openImage(inputDir + image.group()) # open Image
# read calibration for later calculation of distances in um.
calibration = imp.getCalibration()
pxWidth = calibration.pixelWidth
print "px depth", calibration.pixelDepth
timeInterval = round(calibration.frameInterval)
#start measurement
splitCh = CS.split(imp) # split channels
#try:
ATA().segAndMeasure(splitCh[0], splitCh[1]) # perform segmentation and measurement
#else: go to next element in moFileList
# move image to "segProblem" folder
# continue
WindowManager.getImage("binProjMerged").close()
WindowManager.getImage("DUP_C1-"+image.group()).close()
WindowManager.getImage("DUP_C2-"+image.group()).close()
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 runOneFile(fullFilePath):
global gNumChannels
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 = ''
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)
#this is too much work for ScanImage4
#try and guess channels if it is a scanimage file
#scanImage3 = string.find(infoStr, 'scanimage') != -1
#scanimage4 = find(infoStr, 'scanimage.SI4.channelSave = ')
#print 'scanimage4:', scanimage4
#
# 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()
infoStr += 'bAlignBatch6=' + time.strftime("%Y%m%d") + '\n'
#
# crop
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
imp.setRoi(roi)
#time.sleep(1)
#bPrintLog('cropping', 1)
IJ.run('Crop')
infoStr += 'cropping=' + str(gCropLeft) + ',' + str(gCropTop) + ',' + str(gCropWidth) + ',' + str(gCropHeight) + '\n'
#
# remove negative (<0) pixel values
#ip = imp.getProcessor()
#pixels = ip.getPixels() # this returns a reference (not a copy)
#for i in xrange(len(pixels)):
# if pixels[i] < 0:
# pixels[i] = 0
# or this, for each pixel 'x', if x<0 then 0 else x
#pixels = map(lambda x: 0 if x<0 else x, pixels)
#set our new values (without pixels <0) back into original (I hope this handles stacks and channels???)
#ip.setPixels(pixels)
#
# remove calibration
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)
#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)
#msgStr = 'adding calCoeff[0]='+str(calCoeff[0]) + ' to stack.'
#bPrintLog(msgStr, 2)
#subArgVal = 'value=%s stack' % (calCoeff[0],)
#IJ.run('Add...', subArgVal)
# 20150701, 'shift everybody over by linear calibration intercept calCoeff[0]'
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 = 32768 - 128
msgStr = 'Subtracting a magic number '+str(magicNumber) + ' from stack.'
bPrintLog(msgStr, 2)
subArgVal = 'value=%s stack' % (origMin,)
IJ.run('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)
#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()
#append calibration to info string
infoStr += 'calibCoeff_a = ' + str(calCoeff[0]) + '\n'
infoStr += 'calibCoeff_b = ' + str(calCoeff[1]) + '\n'
infoStr += 'origMin = ' + str(origMin) + '\n'
infoStr += 'origMax = ' + str(origMax) + '\n'
#
# set up
if gNumChannels == 1:
impWinStr = imp.getTitle() #use this when only one channel
impWin = WindowManager.getWindow(impWinStr) #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
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 imp.getNSlices()>1:
infoStr += 'AlignOnChannel=1' + '\n'
#snap to middle slice
if gAlignOnMiddleSlice:
middleSlice = int(math.floor(imp.getNSlices() / 2)) #int() is necc., python is f*****g picky
else:
middleSlice = gAlignOnThisSlice
imp.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'
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 += 'AlignOnSlice=' + 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)
#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
ch1Imp.setProperty("Info", infoStr);
#bPrintLog('Saving:' + ch1File, 1)
bSaveStack(ch1Imp, ch1File)
#max project
bSaveZProject(ch1Imp, destMaxFolder, shortName+'_ch1')
#ch2
ch2Imp.setProperty("Info", infoStr);
#bPrintLog('Saving:' + ch2File, 1)
bSaveStack(ch2Imp, ch2File)
#max project
bSaveZProject(ch2Imp, 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:' + ch1WinStr, 1)
IJ.selectWindow(ch1WinStr)
#IJ.run('resetMinAndMax()')
#ch1Imp.resetStack()
#ch1Imp.resetDisplayRange()
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch1.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(ch1Imp, impFile)
#max project
bSaveZProject(ch1Imp, eightBitMaxFolder, shortName+'_ch1')
#
bPrintLog('Converting to 8-bit:' + ch2WinStr, 1)
IJ.selectWindow(ch2WinStr)
#IJ.run('resetMinAndMax()')
IJ.run("8-bit")
impFile = eightBitFolder + shortName + '_ch2.tif'
bPrintLog('Saving 8-bit:' + impFile, 2)
bSaveStack(ch2Imp, impFile)
#max project
bSaveZProject(ch2Imp, eightBitMaxFolder, shortName+'_ch2')
#
# close original window
imp.changes = 0
imp.close()
#
# close ch1/ch2
if 1 and gNumChannels == 2:
ch1Imp.changes = 0
ch1Imp.close()
ch2Imp.changes = 0
ch2Imp.close()
bPrintLog(time.strftime("%H:%M:%S") + ' finished runOneFile(): ' + fullFilePath, 1)
