Beispiel #1
0
def convert32to16(fullFilePath):  #overwrite the file!!!
    if not os.path.isfile(fullFilePath):
        bPrintLog(
            '\nERROR: runOneFile() did not find file: ' + fullFilePath + '\n',
            0)
        return 0
    elif not fullFilePath.endswith('tif'):
        msg = fullFilePath + " is not a deconvoluted tif file"
        bPrintLog(msg, 2)
    else:
        bPrintLog(
            time.strftime("%H:%M:%S") +
            ' starting runOneFile()for overwrite: ' + fullFilePath, 1)

        #open and overwrite the deconvoluted file as 16 bit
        imp = Opener().openImage(fullFilePath)  #deconvoluted file
        if imp.getBitDepth() == 32:
            imp.show()
            msgStr = "Converting 32 to 16-bit..."
            bPrintLog(msgStr, 3)
            IJ.run("16-bit")

            msgStr = "Overwriting 32-bit with 16-bit File in" + fullFilePath
            bPrintLog(msgStr, 3)
            IJ.run(imp, "Save", "")  #save as replace without asking

        imp.close()
def convert32to16(fullFilePath): #overwrite the file!!!
	if not os.path.isfile(fullFilePath):
		bPrintLog('\nERROR: runOneFile() did not find file: ' + fullFilePath + '\n',0)
		return 0
	elif not fullFilePath.endswith('tif'):
		msg = fullFilePath +" is not a deconvoluted tif file"
		bPrintLog(msg,2)
	else:
		bPrintLog(time.strftime("%H:%M:%S") + ' starting runOneFile()for overwrite: ' + fullFilePath, 1)
	
		#open and overwrite the deconvoluted file as 16 bit
		imp = Opener().openImage(fullFilePath)	#deconvoluted file
		if imp.getBitDepth() == 32:
			imp.show()
			msgStr = "Converting 32 to 16-bit..."
			bPrintLog(msgStr,3)
			IJ.run("16-bit")
			
			msgStr = "Overwriting 32-bit with 16-bit File in" + fullFilePath
			bPrintLog(msgStr,3)
			IJ.run(imp,"Save","") #save as replace without asking
			
		imp.close()
Beispiel #3
0
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 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)
Beispiel #5
0
def runOneFile(fullFilePath):
    global gFileType
    global fileIndex

    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)
    bPrintLog('inputfile is:' + fullFilePath, 1)

    enclosingPath = os.path.dirname(fullFilePath)
    head, tail = os.path.split(
        enclosingPath)  #tail is name of enclosing folder
    enclosingPath += '/'

    # make output folders
    destFolder = enclosingPath + tail + '_short/'
    if not os.path.isdir(destFolder):
        os.makedirs(destFolder)

    # open
    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
    #print 'original infoStr:', infoStr
    if not infoStr:
        infoStr = ''
    infoStr += 'ShortenNames_Version=' + str(gShortenVersion) + '\n'
    infoStr += 'ShortenNames_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)

    #output file name
    outFile = destFolder + tail + '_' + str(fileIndex) + '.tif'
    fileIndex += 1
    bPrintLog('output file is:' + outFile, 1)

    # put original name in header
    infoStr += 'ShortenNames_OriginalFile=' + fullFilePath + '\n'

    # put scanimage header back in
    imp.setProperty("Info", infoStr)

    #save
    bSaveStack(imp, outFile)

    #
    # close original window
    imp.changes = 0
    imp.close()
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)
def runOneFile(fullFilePath):
	global gFileType
	global fileIndex
	
	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)
	bPrintLog('inputfile is:' + fullFilePath, 1)
	
	enclosingPath = os.path.dirname(fullFilePath)
	head, tail = os.path.split(enclosingPath) #tail is name of enclosing folder
	enclosingPath += '/'
	
	# make output folders
	destFolder = enclosingPath + tail + '_short/'
	if not os.path.isdir(destFolder):
		os.makedirs(destFolder)

	# open
	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
	#print 'original infoStr:', infoStr
	if not infoStr:
		infoStr = ''
	infoStr += 'ShortenNames_Version=' + str(gShortenVersion) + '\n'
	infoStr += 'ShortenNames_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)

	#output file name
	outFile = destFolder + tail + '_' + str(fileIndex) + '.tif'
	fileIndex += 1
	bPrintLog('output file is:' + outFile, 1)
	
	# put original name in header
	infoStr += 'ShortenNames_OriginalFile=' + fullFilePath + '\n'
	
	# put scanimage header back in
	imp.setProperty("Info", infoStr);

	#save
	bSaveStack(imp, outFile)

	#
	# close original window
	imp.changes = 0
	imp.close()