UnMicst.py

import numpy as np
from scipy import misc
import os
import logging

import tensorflow.compat.v1 as tf
import shutil
import scipy.io as sio
import fnmatch, glob
import skimage.exposure as sk
import skimage.io
import argparse
import czifile
from nd2reader import ND2Reader
import tifffile
import sys
tf.disable_v2_behavior()
#sys.path.insert(0, 'C:\\Users\\Public\\Documents\\ImageScience')

from toolbox.imtools import *
from toolbox.ftools import *
from toolbox.PartitionOfImage import PI2D
from toolbox import GPUselect

def concat3(lst):
	return tf.concat(lst, 3)


class UNet2D:
	hp = None  # hyper-parameters
	nn = None  # network
	tfTraining = None  # if training or not (to handle batch norm)
	tfData = None  # data placeholder
	Session = None
	DatasetMean = 0
	DatasetStDev = 0

	def setupWithHP(hp):
		UNet2D.setup(hp['imSize'],
					 hp['nChannels'],
					 hp['nClasses'],
					 hp['nOut0'],
					 hp['featMapsFact'],
					 hp['downSampFact'],
					 hp['ks'],
					 hp['nExtraConvs'],
					 hp['stdDev0'],
					 hp['nLayers'],
					 hp['batchSize'])

	def setup(imSize, nChannels, nClasses, nOut0, featMapsFact, downSampFact, kernelSize, nExtraConvs, stdDev0,
			  nDownSampLayers, batchSize):
		UNet2D.hp = {'imSize': imSize,
					 'nClasses': nClasses,
					 'nChannels': nChannels,
					 'nExtraConvs': nExtraConvs,
					 'nLayers': nDownSampLayers,
					 'featMapsFact': featMapsFact,
					 'downSampFact': downSampFact,
					 'ks': kernelSize,
					 'nOut0': nOut0,
					 'stdDev0': stdDev0,
					 'batchSize': batchSize}

		nOutX = [UNet2D.hp['nChannels'], UNet2D.hp['nOut0']]
		dsfX = []
		for i in range(UNet2D.hp['nLayers']):
			nOutX.append(nOutX[-1] * UNet2D.hp['featMapsFact'])
			dsfX.append(UNet2D.hp['downSampFact'])

		# --------------------------------------------------
		# downsampling layer
		# --------------------------------------------------

		with tf.name_scope('placeholders'):
			UNet2D.tfTraining = tf.placeholder(tf.bool, name='training')
			UNet2D.tfData = tf.placeholder("float", shape=[None, UNet2D.hp['imSize'], UNet2D.hp['imSize'],
														   UNet2D.hp['nChannels']], name='data')

		def down_samp_layer(data, index):
			with tf.name_scope('ld%d' % index):
				ldXWeights1 = tf.Variable(
					tf.truncated_normal([UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[index], nOutX[index + 1]],
										stddev=stdDev0), name='kernel1')
				ldXWeightsExtra = []
				for i in range(nExtraConvs):
					ldXWeightsExtra.append(tf.Variable(
						tf.truncated_normal([UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[index + 1], nOutX[index + 1]],
											stddev=stdDev0), name='kernelExtra%d' % i))

				c00 = tf.nn.conv2d(data, ldXWeights1, strides=[1, 1, 1, 1], padding='SAME')
				for i in range(nExtraConvs):
					c00 = tf.nn.conv2d(tf.nn.relu(c00), ldXWeightsExtra[i], strides=[1, 1, 1, 1], padding='SAME')

				ldXWeightsShortcut = tf.Variable(
					tf.truncated_normal([1, 1, nOutX[index], nOutX[index + 1]], stddev=stdDev0), name='shortcutWeights')
				shortcut = tf.nn.conv2d(data, ldXWeightsShortcut, strides=[1, 1, 1, 1], padding='SAME')

				bn = tf.layers.batch_normalization(tf.nn.relu(c00 + shortcut), training=UNet2D.tfTraining)

				return tf.nn.max_pool(bn, ksize=[1, dsfX[index], dsfX[index], 1],
									  strides=[1, dsfX[index], dsfX[index], 1], padding='SAME', name='maxpool')

		# --------------------------------------------------
		# bottom layer
		# --------------------------------------------------

		with tf.name_scope('lb'):
			lbWeights1 = tf.Variable(tf.truncated_normal(
				[UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[UNet2D.hp['nLayers']], nOutX[UNet2D.hp['nLayers'] + 1]],
				stddev=stdDev0), name='kernel1')

			def lb(hidden):
				return tf.nn.relu(tf.nn.conv2d(hidden, lbWeights1, strides=[1, 1, 1, 1], padding='SAME'), name='conv')

		# --------------------------------------------------
		# downsampling
		# --------------------------------------------------

		with tf.name_scope('downsampling'):
			dsX = []
			dsX.append(UNet2D.tfData)

			for i in range(UNet2D.hp['nLayers']):
				dsX.append(down_samp_layer(dsX[i], i))

			b = lb(dsX[UNet2D.hp['nLayers']])

		# --------------------------------------------------
		# upsampling layer
		# --------------------------------------------------

		def up_samp_layer(data, index):
			with tf.name_scope('lu%d' % index):
				luXWeights1 = tf.Variable(
					tf.truncated_normal([UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[index + 1], nOutX[index + 2]],
										stddev=stdDev0), name='kernel1')
				luXWeights2 = tf.Variable(tf.truncated_normal(
					[UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[index] + nOutX[index + 1], nOutX[index + 1]],
					stddev=stdDev0), name='kernel2')
				luXWeightsExtra = []
				for i in range(nExtraConvs):
					luXWeightsExtra.append(tf.Variable(
						tf.truncated_normal([UNet2D.hp['ks'], UNet2D.hp['ks'], nOutX[index + 1], nOutX[index + 1]],
											stddev=stdDev0), name='kernel2Extra%d' % i))

				outSize = UNet2D.hp['imSize']
				for i in range(index):
					outSize /= dsfX[i]
				outSize = int(outSize)

				outputShape = [UNet2D.hp['batchSize'], outSize, outSize, nOutX[index + 1]]
				us = tf.nn.relu(
					tf.nn.conv2d_transpose(data, luXWeights1, outputShape, strides=[1, dsfX[index], dsfX[index], 1],
										   padding='SAME'), name='conv1')
				cc = concat3([dsX[index], us])
				cv = tf.nn.relu(tf.nn.conv2d(cc, luXWeights2, strides=[1, 1, 1, 1], padding='SAME'), name='conv2')
				for i in range(nExtraConvs):
					cv = tf.nn.relu(tf.nn.conv2d(cv, luXWeightsExtra[i], strides=[1, 1, 1, 1], padding='SAME'),
									name='conv2Extra%d' % i)
				return cv

		# --------------------------------------------------
		# final (top) layer
		# --------------------------------------------------

		with tf.name_scope('lt'):
			ltWeights1 = tf.Variable(tf.truncated_normal([1, 1, nOutX[1], nClasses], stddev=stdDev0), name='kernel')

			def lt(hidden):
				return tf.nn.conv2d(hidden, ltWeights1, strides=[1, 1, 1, 1], padding='SAME', name='conv')

		# --------------------------------------------------
		# upsampling
		# --------------------------------------------------

		with tf.name_scope('upsampling'):
			usX = []
			usX.append(b)

			for i in range(UNet2D.hp['nLayers']):
				usX.append(up_samp_layer(usX[i], UNet2D.hp['nLayers'] - 1 - i))

			t = lt(usX[UNet2D.hp['nLayers']])

		sm = tf.nn.softmax(t, -1)
		UNet2D.nn = sm

	def train(imPath, logPath, modelPath, pmPath, nTrain, nValid, nTest, restoreVariables, nSteps, gpuIndex,
			  testPMIndex):
		os.environ['CUDA_VISIBLE_DEVICES'] = '%d' % gpuIndex

		outLogPath = logPath
		trainWriterPath = pathjoin(logPath, 'Train')
		validWriterPath = pathjoin(logPath, 'Valid')
		outModelPath = pathjoin(modelPath, 'model.ckpt')
		outPMPath = pmPath

		batchSize = UNet2D.hp['batchSize']
		imSize = UNet2D.hp['imSize']
		nChannels = UNet2D.hp['nChannels']
		nClasses = UNet2D.hp['nClasses']

		# --------------------------------------------------
		# data
		# --------------------------------------------------

		Train = np.zeros((nTrain, imSize, imSize, nChannels))
		Valid = np.zeros((nValid, imSize, imSize, nChannels))
		Test = np.zeros((nTest, imSize, imSize, nChannels))
		LTrain = np.zeros((nTrain, imSize, imSize, nClasses))
		LValid = np.zeros((nValid, imSize, imSize, nClasses))
		LTest = np.zeros((nTest, imSize, imSize, nClasses))

		print('loading data, computing mean / st dev')
		if not os.path.exists(modelPath):
			os.makedirs(modelPath)
		if restoreVariables:
			datasetMean = loadData(pathjoin(modelPath, 'datasetMean.data'))
			datasetStDev = loadData(pathjoin(modelPath, 'datasetStDev.data'))
		else:
			datasetMean = 0
			datasetStDev = 0
			for iSample in range(nTrain + nValid + nTest):
				I = im2double(tifread('%s/I%05d_Img.tif' % (imPath, iSample)))
				datasetMean += np.mean(I)
				datasetStDev += np.std(I)
			datasetMean /= (nTrain + nValid + nTest)
			datasetStDev /= (nTrain + nValid + nTest)
			saveData(datasetMean, pathjoin(modelPath, 'datasetMean.data'))
			saveData(datasetStDev, pathjoin(modelPath, 'datasetStDev.data'))

		perm = np.arange(nTrain + nValid + nTest)
		np.random.shuffle(perm)

		for iSample in range(0, nTrain):
			path = '%s/I%05d_Img.tif' % (imPath, perm[iSample])
			im = im2double(tifread(path))
			Train[iSample, :, :, 0] = (im - datasetMean) / datasetStDev
			path = '%s/I%05d_Ant.tif' % (imPath, perm[iSample])
			im = tifread(path)
			for i in range(nClasses):
				LTrain[iSample, :, :, i] = (im == i + 1)

		for iSample in range(0, nValid):
			path = '%s/I%05d_Img.tif' % (imPath, perm[nTrain + iSample])
			im = im2double(tifread(path))
			Valid[iSample, :, :, 0] = (im - datasetMean) / datasetStDev
			path = '%s/I%05d_Ant.tif' % (imPath, perm[nTrain + iSample])
			im = tifread(path)
			for i in range(nClasses):
				LValid[iSample, :, :, i] = (im == i + 1)

		for iSample in range(0, nTest):
			path = '%s/I%05d_Img.tif' % (imPath, perm[nTrain + nValid + iSample])
			im = im2double(tifread(path))
			Test[iSample, :, :, 0] = (im - datasetMean) / datasetStDev
			path = '%s/I%05d_Ant.tif' % (imPath, perm[nTrain + nValid + iSample])
			im = tifread(path)
			for i in range(nClasses):
				LTest[iSample, :, :, i] = (im == i + 1)

		# --------------------------------------------------
		# optimization
		# --------------------------------------------------

		tfLabels = tf.placeholder("float", shape=[None, imSize, imSize, nClasses], name='labels')

		globalStep = tf.Variable(0, trainable=False)
		learningRate0 = 0.01
		decaySteps = 1000
		decayRate = 0.95
		learningRate = tf.train.exponential_decay(learningRate0, globalStep, decaySteps, decayRate, staircase=True)

		with tf.name_scope('optim'):
			loss = tf.reduce_mean(-tf.reduce_sum(tf.multiply(tfLabels, tf.log(UNet2D.nn)), 3))
			updateOps = tf.get_collection(tf.GraphKeys.UPDATE_OPS)
			# optimizer = tf.train.MomentumOptimizer(1e-3,0.9)
			optimizer = tf.train.MomentumOptimizer(learningRate, 0.9)
			# optimizer = tf.train.GradientDescentOptimizer(learningRate)
			with tf.control_dependencies(updateOps):
				optOp = optimizer.minimize(loss, global_step=globalStep)

		with tf.name_scope('eval'):
			error = []
			for iClass in range(nClasses):
				labels0 = tf.reshape(tf.to_int32(tf.slice(tfLabels, [0, 0, 0, iClass], [-1, -1, -1, 1])),
									 [batchSize, imSize, imSize])
				predict0 = tf.reshape(tf.to_int32(tf.equal(tf.argmax(UNet2D.nn, 3), iClass)),
									  [batchSize, imSize, imSize])
				correct = tf.multiply(labels0, predict0)
				nCorrect0 = tf.reduce_sum(correct)
				nLabels0 = tf.reduce_sum(labels0)
				error.append(1 - tf.to_float(nCorrect0) / tf.to_float(nLabels0))
			errors = tf.tuple(error)

		# --------------------------------------------------
		# inspection
		# --------------------------------------------------

		with tf.name_scope('scalars'):
			tf.summary.scalar('avg_cross_entropy', loss)
			for iClass in range(nClasses):
				tf.summary.scalar('avg_pixel_error_%d' % iClass, error[iClass])
			tf.summary.scalar('learning_rate', learningRate)
		with tf.name_scope('images'):
			split0 = tf.slice(UNet2D.nn, [0, 0, 0, 0], [-1, -1, -1, 1])
			split1 = tf.slice(UNet2D.nn, [0, 0, 0, 1], [-1, -1, -1, 1])
			if nClasses > 2:
				split2 = tf.slice(UNet2D.nn, [0, 0, 0, 2], [-1, -1, -1, 1])
			tf.summary.image('pm0', split0)
			tf.summary.image('pm1', split1)
			if nClasses > 2:
				tf.summary.image('pm2', split2)
		merged = tf.summary.merge_all()

		# --------------------------------------------------
		# session
		# --------------------------------------------------

		saver = tf.train.Saver()
		sess = tf.Session(config=tf.ConfigProto(
			allow_soft_placement=True))  # config parameter needed to save variables when using GPU

		if os.path.exists(outLogPath):
			shutil.rmtree(outLogPath)
		trainWriter = tf.summary.FileWriter(trainWriterPath, sess.graph)
		validWriter = tf.summary.FileWriter(validWriterPath, sess.graph)

		if restoreVariables:
			saver.restore(sess, outModelPath)
			print("Model restored.")
		else:
			sess.run(tf.global_variables_initializer())

		# --------------------------------------------------
		# train
		# --------------------------------------------------

		batchData = np.zeros((batchSize, imSize, imSize, nChannels))
		batchLabels = np.zeros((batchSize, imSize, imSize, nClasses))
		for i in range(nSteps):
			# train

			perm = np.arange(nTrain)
			np.random.shuffle(perm)

			for j in range(batchSize):
				batchData[j, :, :, :] = Train[perm[j], :, :, :]
				batchLabels[j, :, :, :] = LTrain[perm[j], :, :, :]

			summary, _ = sess.run([merged, optOp],
								  feed_dict={UNet2D.tfData: batchData, tfLabels: batchLabels, UNet2D.tfTraining: 1})
			trainWriter.add_summary(summary, i)

			# validation

			perm = np.arange(nValid)
			np.random.shuffle(perm)

			for j in range(batchSize):
				batchData[j, :, :, :] = Valid[perm[j], :, :, :]
				batchLabels[j, :, :, :] = LValid[perm[j], :, :, :]

			summary, es = sess.run([merged, errors],
								   feed_dict={UNet2D.tfData: batchData, tfLabels: batchLabels, UNet2D.tfTraining: 0})
			validWriter.add_summary(summary, i)

			e = np.mean(es)
			print('step %05d, e: %f' % (i, e))

			if i == 0:
				if restoreVariables:
					lowestError = e
				else:
					lowestError = np.inf

			if np.mod(i, 100) == 0 and e < lowestError:
				lowestError = e
				print("Model saved in file: %s" % saver.save(sess, outModelPath))

		# --------------------------------------------------
		# test
		# --------------------------------------------------

		if not os.path.exists(outPMPath):
			os.makedirs(outPMPath)

		for i in range(nTest):
			j = np.mod(i, batchSize)

			batchData[j, :, :, :] = Test[i, :, :, :]
			batchLabels[j, :, :, :] = LTest[i, :, :, :]

			if j == batchSize - 1 or i == nTest - 1:

				output = sess.run(UNet2D.nn,
								  feed_dict={UNet2D.tfData: batchData, tfLabels: batchLabels, UNet2D.tfTraining: 0})

				for k in range(j + 1):
					pm = output[k, :, :, testPMIndex]
					gt = batchLabels[k, :, :, testPMIndex]
					im = np.sqrt(normalize(batchData[k, :, :, 0]))
					imwrite(np.uint8(255 * np.concatenate((im, np.concatenate((pm, gt), axis=1)), axis=1)),
							'%s/I%05d.png' % (outPMPath, i - j + k + 1))

		# --------------------------------------------------
		# save hyper-parameters, clean-up
		# --------------------------------------------------

		saveData(UNet2D.hp, pathjoin(modelPath, 'hp.data'))

		trainWriter.close()
		validWriter.close()
		sess.close()

	def deploy(imPath, nImages, modelPath, pmPath, gpuIndex, pmIndex):
		os.environ['CUDA_VISIBLE_DEVICES'] = '%d' % gpuIndex

		variablesPath = pathjoin(modelPath, 'model.ckpt')
		outPMPath = pmPath

		hp = loadData(pathjoin(modelPath, 'hp.data'))
		UNet2D.setupWithHP(hp)

		batchSize = UNet2D.hp['batchSize']
		imSize = UNet2D.hp['imSize']
		nChannels = UNet2D.hp['nChannels']
		nClasses = UNet2D.hp['nClasses']

		# --------------------------------------------------
		# data
		# --------------------------------------------------

		Data = np.zeros((nImages, imSize, imSize, nChannels))

		datasetMean = loadData(pathjoin(modelPath, 'datasetMean.data'))
		datasetStDev = loadData(pathjoin(modelPath, 'datasetStDev.data'))

		for iSample in range(0, nImages):
			path = '%s/I%05d_Img.tif' % (imPath, iSample)
			im = im2double(tifread(path))
			Data[iSample, :, :, 0] = (im - datasetMean) / datasetStDev

		# --------------------------------------------------
		# session
		# --------------------------------------------------

		saver = tf.train.Saver()
		sess = tf.Session(config=tf.ConfigProto(
			allow_soft_placement=True))  # config parameter needed to save variables when using GPU

		saver.restore(sess, variablesPath)
		print("Model restored.")

		# --------------------------------------------------
		# deploy
		# --------------------------------------------------

		batchData = np.zeros((batchSize, imSize, imSize, nChannels))

		if not os.path.exists(outPMPath):
			os.makedirs(outPMPath)

		for i in range(nImages):
			print(i, nImages)

			j = np.mod(i, batchSize)

			batchData[j, :, :, :] = Data[i, :, :, :]

			if j == batchSize - 1 or i == nImages - 1:

				output = sess.run(UNet2D.nn, feed_dict={UNet2D.tfData: batchData, UNet2D.tfTraining: 0})

				for k in range(j + 1):
					pm = output[k, :, :, pmIndex]
					im = np.sqrt(normalize(batchData[k, :, :, 0]))
					# imwrite(np.uint8(255*np.concatenate((im,pm),axis=1)),'%s/I%05d.png' % (outPMPath,i-j+k+1))
					imwrite(np.uint8(255 * im), '%s/I%05d_Im.png' % (outPMPath, i - j + k + 1))
					imwrite(np.uint8(255 * pm), '%s/I%05d_PM.png' % (outPMPath, i - j + k + 1))

		# --------------------------------------------------
		# clean-up
		# --------------------------------------------------

		sess.close()

	def singleImageInferenceSetup(modelPath, gpuIndex,mean,std):
		variablesPath = pathjoin(modelPath, 'model.ckpt')

		hp = loadData(pathjoin(modelPath, 'hp.data'))
		UNet2D.setupWithHP(hp)
		if mean ==-1:
			UNet2D.DatasetMean = loadData(pathjoin(modelPath, 'datasetMean.data'))
		else:
			UNet2D.DatasetMean = mean

		if std == -1:
			UNet2D.DatasetStDev = loadData(pathjoin(modelPath, 'datasetStDev.data'))
		else:
			UNet2D.DatasetStDev = std
		print(UNet2D.DatasetMean)
		print(UNet2D.DatasetStDev)

		# --------------------------------------------------
		# session
		# --------------------------------------------------

		saver = tf.train.Saver()
		UNet2D.Session = tf.Session(config=tf.ConfigProto())
			# allow_soft_placement=True))  # config parameter needed to save variables when using GPU

		saver.restore(UNet2D.Session, variablesPath)
		print("Model restored.")

	def singleImageInferenceCleanup():
		UNet2D.Session.close()

	def singleImageInference(image, mode, pmIndex):
		print('Inference...')

		batchSize = UNet2D.hp['batchSize']
		imSize = UNet2D.hp['imSize']
		nChannels = UNet2D.hp['nChannels']

		PI2D.setup(image, imSize, int(imSize / 8), mode)
		PI2D.createOutput(nChannels)

		batchData = np.zeros((batchSize, imSize, imSize, nChannels))
		for i in range(PI2D.NumPatches):
			j = np.mod(i, batchSize)
			batchData[j, :, :, 0] = (PI2D.getPatch(i) - UNet2D.DatasetMean) / UNet2D.DatasetStDev
			if j == batchSize - 1 or i == PI2D.NumPatches - 1:
				output = UNet2D.Session.run(UNet2D.nn, feed_dict={UNet2D.tfData: batchData, UNet2D.tfTraining: 0})
				for k in range(j + 1):
					pm = output[k, :, :, pmIndex]
					PI2D.patchOutput(i - j + k, pm)
			# PI2D.patchOutput(i-j+k,normalize(imgradmag(PI2D.getPatch(i-j+k),1)))

		return PI2D.getValidOutput()


if __name__ == '__main__':
	parser = argparse.ArgumentParser()
	parser.add_argument("imagePath", help="path to the .tif file")
	parser.add_argument("--model",  help="type of model. For example, nuclei vs cytoplasm",default = 'nucleiDAPI')
	parser.add_argument("--outputPath", help="output path of probability map")
	parser.add_argument("--channel", help="channel to perform inference on", type=int, default=0)
	parser.add_argument("--classOrder", help="background, contours, foreground", type = int, nargs = '+', default=-1)
	parser.add_argument("--mean", help="mean intensity of input image. Use -1 to use model", type=float, default=-1)
	parser.add_argument("--std", help="mean standard deviation of input image. Use -1 to use model", type=float, default=-1)
	parser.add_argument("--scalingFactor", help="factor by which to increase/decrease image size by", type=float,
						default=1)
	parser.add_argument("--stackOutput", help="save probability maps as separate files", action='store_true')
	parser.add_argument("--GPU", help="explicitly select GPU", type=int, default = -1)
	parser.add_argument("--outlier",
						help="map percentile intensity to max when rescaling intensity values. Max intensity as default",
						type=float, default=-1)
	parser.add_argument("--verbose", help="display error messages for debugging", action='store_true')
	args = parser.parse_args()

	if args.verbose:
		os.environ['TF_CPP_MIN_LOG_LEVEL'] = '0'
		logging.getLogger('tensorflow').setLevel(logging.DEBUG)
	else:
		os.environ['TF_CPP_MIN_LOG_LEVEL'] = '2'
		logging.getLogger('tensorflow').setLevel(logging.FATAL)

	logPath = ''
	scriptPath = os.path.dirname(os.path.realpath(__file__))
	modelPath = os.path.join(scriptPath, 'models', args.model)
	# modelPath = os.path.join(scriptPath, 'models/cytoplasmINcell')
	# modelPath = os.path.join(scriptPath, 'cytoplasmZeissNikon')
	pmPath = ''

	if os.system('nvidia-smi') == 0:
		if args.GPU == -1:
			print("automatically choosing GPU")
			GPU = GPUselect.pick_gpu_lowest_memory()
		else:
			GPU = args.GPU
		print('Using GPU ' + str(GPU))

	else:
		if sys.platform == 'win32':  # only 1 gpu on windows
			if args.GPU==-1:
				GPU = 0
			else:
				GPU = args.GPU
			print('Using GPU ' + str(GPU))
		else:
			GPU=0
			print('Using CPU')
	os.environ['CUDA_VISIBLE_DEVICES'] = '%d' % GPU
	UNet2D.singleImageInferenceSetup(modelPath, GPU,args.mean,args.std)
	nClass = UNet2D.hp['nClasses']
	imagePath = args.imagePath
	dapiChannel = args.channel
	dsFactor = args.scalingFactor
	parentFolder = os.path.dirname(os.path.dirname(imagePath))
	fileName = os.path.basename(imagePath)
	fileNamePrefix = fileName.split(os.extsep, 1)
	# print(fileName)
	fileType = fileNamePrefix[1]

	if fileType=='ome.tif' or fileType=='ome.tiff' or fileType == 'btf' :
		I = skio.imread(imagePath, img_num=dapiChannel,plugin='tifffile')
	elif fileType == 'tif' :
		I = tifffile.imread(imagePath, key=dapiChannel)
	elif fileType == 'czi':
		with czifile.CziFile(imagePath) as czi:
			image = czi.asarray()
			I = image[0, 0, dapiChannel, 0, 0, :, :, 0]
	elif fileType == 'nd2':
		with ND2Reader(imagePath) as fullStack:
			I = fullStack[dapiChannel]
	if I.dtype == 'float32':
		I=np.uint16(I)
	if args.classOrder == -1:
		args.classOrder = range(nClass)

	rawI = I
	# print(type(I))
	hsize = int((float(I.shape[0]) * float(dsFactor)))
	vsize = int((float(I.shape[1]) * float(dsFactor)))
	I = resize(I, (hsize, vsize))
	if args.outlier == -1:
		maxLimit = np.max(I)
	else:
		maxLimit = np.percentile(I, args.outlier)
	I = im2double(sk.rescale_intensity(I, in_range=(np.min(I), maxLimit), out_range=(0, 0.983)))
	rawI = im2double(rawI) / np.max(im2double(rawI))
	if not args.outputPath:
		args.outputPath = parentFolder + '//probability_maps'

	if not os.path.exists(args.outputPath):
		os.makedirs(args.outputPath)
	os.makedirs(args.outputPath + '//qc', exist_ok=True)

	append_kwargs = {
		'bigtiff': True,
		'metadata': None,
		'append': True,
	}
	save_kwargs = {
		'bigtiff': True,
		'metadata': None,
		'append': False,
	}
	if args.stackOutput:
		slice=0
		for iClass in args.classOrder[::-1]:
			PM = np.uint8(255*UNet2D.singleImageInference(I, 'accumulate', iClass)) # backwards in order to align with ilastik...
			PM = resize(PM, (rawI.shape[0], rawI.shape[1]))
			if slice==0:
				skimage.io.imsave(args.outputPath + '//' + fileNamePrefix[0] + '_Probabilities_' + str(dapiChannel+1) + '.tif', np.uint8(255 * PM),**save_kwargs)
			else:
				skimage.io.imsave(args.outputPath + '//' + fileNamePrefix[0] + '_Probabilities_' + str(dapiChannel+1) + '.tif',np.uint8(255 * PM),**append_kwargs)
			if slice==1:
				save_kwargs['append'] = False
				skimage.io.imsave(args.outputPath + '//qc//' + fileNamePrefix[0] + '_Preview_' + str(dapiChannel+1) + '.tif',	np.uint8(255 * PM), **save_kwargs)
				skimage.io.imsave(args.outputPath + '//qc//' + fileNamePrefix[0] + '_Preview_' + str(dapiChannel+1) + '.tif', np.uint8(255 * rawI), **append_kwargs)
			slice = slice + 1

	else:
		contours = np.uint8(255*UNet2D.singleImageInference(I, 'accumulate', args.classOrder[1]))
		hsize = int((float(I.shape[0]) * float(1 / dsFactor)))
		vsize = int((float(I.shape[1]) * float(1 / dsFactor)))
		contours = resize(contours, (rawI.shape[0], rawI.shape[1]))
		skimage.io.imsave(args.outputPath + '//' + fileNamePrefix[0] + '_ContoursPM_' + str(dapiChannel+1) + '.tif',np.uint8(255 * contours),**save_kwargs)
		skimage.io.imsave(args.outputPath + '//' + fileNamePrefix[0] + '_ContoursPM_' + str(dapiChannel+1) + '.tif',np.uint8(255 * rawI), **append_kwargs)
		del contours
		nuclei = np.uint8(255*UNet2D.singleImageInference(I, 'accumulate', args.classOrder[2]))
		nuclei = resize(nuclei, (rawI.shape[0], rawI.shape[1]))
		skimage.io.imsave(args.outputPath + '//' + fileNamePrefix[0] + '_NucleiPM_' + str(dapiChannel+1) + '.tif',np.uint8(255 * nuclei), **save_kwargs)
		del nuclei
	UNet2D.singleImageInferenceCleanup()