def keep_blobs_bigger_than(imp, min_size_pix=100):
"""remove all blobs other than the largest by area"""
imp.killRoi()
rt = ResultsTable()
if "Size_filtered_" in imp.getTitle():
title_addition = ""
else:
title_addition = "Size_filtered_"
out_imp = IJ.createImage("{}{}".format(title_addition, imp.getTitle()),
imp.getWidth(), imp.getHeight(), 1, 8)
#out_imp.show()
IJ.run(out_imp, "Select All", "")
IJ.run(out_imp, "Set...", "value=0 slice")
mxsz = imp.width * imp.height
roim = RoiManager(False)
pa = ParticleAnalyzer(ParticleAnalyzer.ADD_TO_MANAGER,
ParticleAnalyzer.AREA | ParticleAnalyzer.SLICE, rt,
min_size_pix, mxsz)
pa.setRoiManager(roim)
roim.reset()
rt.reset()
pa.analyze(imp)
if roim.getCount() > 0:
rt_areas = rt.getColumn(rt.getColumnIndex("Area")).tolist()
# print("Number of cells identified: {}".format(len(rt_areas)))
for idx in range(len(rt_areas)):
roim.select(out_imp, idx)
IJ.run(out_imp, "Set...", "value=255 slice")
roim.reset()
roim.close()
imp.changes = False
imp.close()
return out_imp
def save_all(srcDir, dstDir, filename, localfile, keepDirectories, imageID):
saveDir = currentDir.replace(srcDir, dstDir) if keepDirectories else dstDir
if not os.path.exists(saveDir):
os.makedirs(saveDir)
print "Saving to", saveDir
IJ.selectWindow("Results for PA ")
IJ.saveAs("Text", ""+saveDir+"\\"+localfile+".xls")
IJ.selectWindow(imageID);
imp=IJ.getImage()
IJ.run("Duplicate...", "duplicate channels="+str(y)+"") #brightfield channel
IJ.run("Z Project...", "projection=[Sum Slices]")
rm= RoiManager(True)
rm.runCommand("Show All without labels")
rm.runCommand("Set Color", "red")
rm.runCommand("Set Line Width", str(4))
imp=IJ.getImage()
IJ.saveAs(imp,"Tiff", ""+saveDir+"\\"+localfile+"_OV.tif")
rm.runCommand("Show None")
IJ.saveAs(imp,"Tiff", ""+saveDir+"\\"+localfile+".tif")
IJ.selectWindow("IMAGE")
imp=IJ.getImage()
IJ.saveAs(imp, "Tiff", os.path.join(saveDir, filename))
ROInumber = rm.getCount()
if(ROInumber>0):
RoiManager("Delete")
def perform_manual_qc(imp, rois, important_channel=1):
"""given cell rois generated by automatic methods, allow user to delete/add/redraw as appropriate"""
for ch in range(imp.getNChannels()):
imp.setC(ch + 1)
sat_frac = 0.99 if (ch + 1) == important_channel else 0.01
IJ.run(imp, "Enhance Contrast", "saturated={}".format(sat_frac))
imp.setC(important_channel)
IJ.setTool("freehand")
proceed = False
roim = RoiManager()
roim.runCommand("Show all with labels")
for roi in rois:
roim.addRoi(roi)
auto_rois_only = rois
while not proceed:
dialog = NonBlockingGenericDialog("Perform manual segmentation")
dialog.setOKLabel("Proceed to next image...")
dialog.addMessage("Perform manual correction of segmentation: ")
dialog.addMessage(
"Draw around cells and add to the region of interest manager (Ctrl+T). "
)
dialog.addMessage("Delete and redraw cells as appropriate. ")
dialog.addMessage(
"Then press \"proceed to next image\" when all cells have been added. "
)
dialog.showDialog()
if dialog.wasCanceled():
print("Manual segmentation canceled")
return auto_rois_only
elif dialog.wasOKed():
if roim.getCount() == 0:
rois = []
confirm_dialog = GenericDialog("Continue?")
confirm_dialog.addMessage(
"No rois selected in this FOV. Are you sure you want to proceed?"
)
confirm_dialog.setOKLabel("Yes, proceed")
confirm_dialog.setCancelLabel("No, not yet")
confirm_dialog.showDialog()
if confirm_dialog.wasOKed():
proceed = True
else:
rois = roim.getRoisAsArray()
proceed = True
roim.reset()
roim.close()
for ch in range(imp.getNChannels()):
imp.setC(ch + 1)
IJ.run(imp, "Enhance Contrast", "saturated={}".format(0.35))
imp.setC(important_channel)
return rois
#Checking if savepath exists
# try:
# os.makedirs(savepath)
# except OSError as e:
# if e.errno != errno.EEXIST:
# raise
Nuclei = Segment_Nuclei(imp1, nucleus_channel)
IJ.run(
Nuclei, "Analyze Particles...",
"size=5000-Infinity pixel circularity=0.3-1.00 add exclude stack")
IJ.run("Tile", "")
time.sleep(0.2)
name1 = imp1.getTitle()
nuclei_count = rm.getCount()
if rm.getCount() != 0:
rois = rm.getIndexes()
rm.setSelectedIndexes(rois)
if rm.getCount() > 1:
rm.runCommand("Combine")
rm.runCommand("Delete")
rm.runCommand("Add")
rois1 = rm.getIndexes()
for roi in rois1:
rm.select(imp1, roi)
imp1.setC(1)
IJ.run(imp1, "Draw", "slice")
IJ.selectWindow(name1)
def run():
zk = ZeissKeys()
msg = Message()
timestart = time.clock()
channel = 1
#size in um2
minArea = 20
maxArea = 200
maxMetaArea = 70
minMetaAR = 1.8
method = "Triangle"
print "Enter run"
detectionNucleus = [minArea, maxArea]
detectionMetaphase = [minArea, maxMetaArea]
filepath = ""
try:
filepath = getArgument()
image = IJ.openImage(filepath) #arguments provided by runMacro
except NameError:
try:
filepath = newImagePath #argument provided by the macroRunner
except NameError: #no file name specified
errMsg = "Fiji error segmentNuclei.py: no filepath has been passed to the macro"
exitWithError(errMsg)
od = OpenDialog("Choose image file", None)
if od.getFileName() == None:
return
filepath = os.path.join(od.getDirectory(), od.getFileName())
# A roimanager is recquired
roim = RoiManager.getInstance()
if roim == None:
roim = RoiManager()
#test a last time
if roim == None:
print 'Fiji error segmentNuclei.py: no RoiManager!'
exitWithError('Fiji error segmentNuclei.py: no RoiManager!')
try:
IJ.run("Close all forced")
IJ.run("Clear Results")
IJ.log("\\Clear")
image = IJ.openImage(filepath)
IJ.log("File Path " + filepath)
try:
image.hide()
except:
image = IJ.getImage()
image.hide()
image.show()
if image is None:
exitWithError("Fiji failed to open image")
return 0
#convert um2 to pixels
pixelSize = image.getCalibration().pixelWidth
detectionNucleus = [area/pixelSize/pixelSize for area in detectionNucleus]
detectionMetaphase = [area/pixelSize/pixelSize for area in detectionMetaphase]
detectionMetaphase.append(minMetaAR)
title = image.getTitle()
imageC = None
#split colors if needed
if image.getNChannels() > 1:
chSp = ChannelSplitter()
imageC = chSp.split(image)
imageSeg = imageC[channel-1].duplicate()
else:
imageSeg = image.duplicate()
imageSeg.setTitle("SEG_"+title)
sp = segment_Particles()
imageSeg = sp.preprocessImage(imageSeg, "MAX", "C1", 2, 2)
imageSeg = sp.segmentParticles(imageSeg, method, detectionNucleus[0], detectionNucleus[1], 2, 2)
roim = RoiManager.getInstance()
if roim == None:
print 'Fiji error segmentNuclei.py: no RoiManager!'
exitWithError('Fiji error segmentNuclei.py: no RoiManager!')
#compute central object
if roim.getCount() > 0:
centPart, msg.position = centerParticle(imageSeg, roim)
msg.position = msg.position + (focusParticle(image, roim, centPart),)
writePositionToRegistry(msg.position)
else:
IJ.run("Read Write Windows Registry", "action=write location=[HKCU\\"+zk.regkey+"] key="+zk.subkey_codemic+" value=["+msg.CODE_NOTHING+"] windows=REG_SZ")
return 0
#create output image
if imageC is None:
impOut = createOutputImg(image, roim, centPart, 0)
else:
impOut = createOutputImg(imageC[channel-1], roim, centPart, 0)
impOut.show()
#save outut image
#saveOutputImg(impOut, filepath, 'Analyzed')
#check for roi properties to identofy metaphase
metaphase = isSpecialParticle(image, roim, centPart,detectionNucleus, detectionMetaphase,pixelSize, msg.position[2])
if os.path.basename(filepath).startswith('TR1_') or '_TR1_' in os.path.basename(filepath):
metaphase = 0
if metaphase:
IJ.run("Read Write Windows Registry", "action=write location=[HKCU\\"+zk.regkey+"] key="+zk.subkey_codemic+" value=["+msg.CODE_TRIGGER1+"] windows=REG_SZ")
else:
IJ.run("Read Write Windows Registry", "action=write location=[HKCU\\"+zk.regkey+"] key="+zk.subkey_codemic+" value=["+msg.CODE_FOCUS+"] windows=REG_SZ")
IJ.log("time focus " + str(time.clock() - timestart))
IJ.run("Collect Garbage")
#IJ.run("Close all forced")
#IJ.run("Clear Results")
except BaseException, err:
# ImageJ Python (Jython) script by Theresa Swayne, Columbia University, 2016
# Purpose: Select random ROIs from a grid produced by EPFL VSI Reader
# to start with, open your VSI and select a rectangular area of tissue, then click Grid.
# if you have a lot of ROIs that have no tissue, delete these before running
from ij import IJ
from ij.plugin.frame import RoiManager
from ij.gui import Roi
rm = RoiManager.getInstance()
if not rm:
rm = RoiManager()
# number of squares in the grid = number of values to choose from
gridSquares = rm.getCount()
# number of fields you need (must be <= gridSquares)
fieldsNeeded = 6
# get path for temp file
import os
from ij.io import DirectoryChooser
dc = DirectoryChooser("Pick folder for saving ROI temp file")
folder = dc.getDirectory()
# make a list of random integers
# ROI manager indices start with 0
def __displayCells(self, nameimage, methodeleon=False):
"""
Displays all the ROIs of the cells with different colors
"""
# we define a list of colors that will be used.
colors = []
ncells= len(self.__dict[nameimage])
if ncells > 0 :
step=200/ncells
if step<1 : step=1
for i in range(ncells) :
r = random.randrange(5,205,step)
g = random.randrange(10,210,step)
b = random.randrange(30,230,step)
#r = int(0+i*step)
#g = random.randrange(10, 190, 30)
#b = int(250-i*step)
colors.append(Color(r, g, b))
else : colors=[Color.blue, Color.green, Color.magenta, Color.orange, Color.yellow]
tempcolors=list(colors)
# we try to have random and different colors for each cell.
for cellname in self.__dict[nameimage].keys() :
if len(tempcolors)>0 :
self.__dict[nameimage][cellname].setColor(tempcolors.pop(0))
else :
tempcolors=list(colors)
self.__dict[nameimage][cellname].setColor(tempcolors.pop(0))
self.__SaveCells(nameimage)
rm = RoiManager.getInstance()
if (rm==None): rm = RoiManager()
rm.runCommand("reset")
# if the user wants to save files, .zip for the ROIs are saved.
#if self.__optionSave == True :
#os.mkdir(self.__pathdir+"ROIs/", mode=0777)
os.makedirs(self.__pathdir+"ROIs/", mode=0777)
tempimp = IJ.createImage("tempimp", "8-bit Black", self.__dictImages[nameimage].getWidth(), self.__dictImages[nameimage].getHeight(), 1)
tempimp.show()
for cellname in self.__dict[nameimage].keys() :
for numslice in range(self.__dictImages[nameimage].getImageStackSize()) :
r = self.__dict[nameimage][cellname].getRoi(numslice)
try :
name=r.getName()
except AttributeError : continue
else :
s = "%04i" % (numslice+1)
#name=s+"-"+name.split("-", 1)[1]
name=s+"-cell"+name.split("cell")[1]
r.setName(name)
try :
rm.addRoi(r)
rname=rm.getName(rm.getCount()-1)
#rm.select(self.__dictImages[nameimage], rm.getCount()-1)
rm.select(tempimp, rm.getCount()-1)
rm.runCommand("Rename", name)
except TypeError : continue
#if isinstance(self.__dict[nameimage][cellname].getRoi(numslice),Roi) == True :
# s = "%04i" % (numslice)
# #rm.add(self.__dictImages[nameimage], self.__dict[nameimage][cellname].getRoi(numslice) , numslice)
# name=self.__dict[nameimage][cellname].getRoi(numslice).getName()
# name=s+name
# self.__dict[nameimage][cellname].getRoi(numslice).setName(name)
# rm.addRoi(self.__dict[nameimage][cellname].getRoi(numslice))
rm.runCommand("Save", self.__pathdir+"ROIs/"+cellname+".zip")
rm.runCommand("reset")
tempimp.close()
def __settings(self, imgName) :
"""
Lets the user to choose different measures to make, and displays it following the choice of the user.
"""
try : dico=self.__dictCells[imgName]
except KeyError :
try : dico=self.__dictCells[imgName[:-4]]
except KeyError : return False
else : imgName=imgName[:-4]
dico=self.__dictCells[imgName]
for cellname in dico.keys() :
self.__dictMeasures[dico[cellname]]={}
# Represents the datas on a diagram
def diagrambuttonPressed(event) :
IJ.showMessage("Push 'Auto' button each time you want to see the diagram")
x1=10
y1=20
x2=100
y2=50
x3=60
y3=30
xr=10
yr=20
wr=20
hr=20
rect=Rectangle(xr,yr,wr,hr)
#img=IJ.getImage()
#nbslices=self.__img.getImageStackSize()
nbslices=self.__maxLife
IJ.run("Hyperstack...", "title=Diagram type=32-bit display=Color width="+str(x2+(nbslices+1)*x3)+" height="+str(y2+y3*len(dico))+" channels=1 slices="+str(len(self.__measures))+" frames=1")
im=IJ.getImage()
ip=im.getProcessor()
for i in range(len(self.__measures)) :
indiceligne=0
maxvalue=0
minvalue=1000000
im.setPosition(1,i+1,1)
for cellname in self.__listcellname :
indiceligne+=1
for indicecolonne in range(1,nbslices+1):
rect.setLocation(x2+indicecolonne*x3+int(x3/6),(y1+indiceligne*y3-int(y3/2)))
# we create at the first iteration a dictionary with the rectangles (for a future use)
if i==0 :
self.__gridrectangle[(indiceligne,indicecolonne)]=Rectangle(rect)
im.setRoi(rect)
ipr=im.getProcessor()
# we find the min and max values of the datas for a measure given.
if self.__dictMeasures[dico[cellname]][self.__measures[i]][indicecolonne-1]>maxvalue :
maxvalue=self.__dictMeasures[dico[cellname]][self.__measures[i]][indicecolonne-1]
if self.__dictMeasures[dico[cellname]][self.__measures[i]][indicecolonne-1]<minvalue :
minvalue=self.__dictMeasures[dico[cellname]][self.__measures[i]][indicecolonne-1]
# we fill the rectangle with the value of the measure
ipr.setValue(self.__dictMeasures[dico[cellname]][self.__measures[i]][indicecolonne-1])
ipr.fill()
# we write the names and the n of slices on the image with the maxvalue.
ip.setValue(maxvalue)
ip.moveTo(x1,y1)
ip.drawString(self.__measures[i])
for j in range(1,nbslices+1) :
ip.moveTo(x2+j*x3,y1)
ip.drawString("Slice "+str(j))
j=0
for cellname in self.__listcellname :
ip.moveTo(x1,y2+j*y3)
ip.drawString(cellname)
j+=1
im.killRoi()
im=IJ.run(im,"Fire","")
IJ.run("Brightness/Contrast...", "")
#im.setMinAndMax(minvalue,maxvalue)
#im.updateImage()
#we add a mouse listener in order to be able to show the roi corresponding to a rectangle when the user clicks on it.
listener = ML()
listener.name=imgName
for imp in map(WindowManager.getImage, WindowManager.getIDList()):
if imp.getTitle().startswith("Diagram") :
win = imp.getWindow()
if win is None:
continue
win.getCanvas().addMouseListener(listener)
# Represents the datas on a series of graphs.
def graphbuttonPressed(event) :
colors=[]
#img=IJ.getImage()
#nbslices=self.__img.getImageStackSize()
nbslices=self.__maxLife
acell=dico.values()[0]
if self.__useTime :
x = acell.getListTimes()
namex="Time sec"
else :
x = range(1,nbslices+1)
namex = "Frame"
maxx=max(x)
minx=min(x)
#x=[i for i in range(1,nbslices+1)]
font=Font("new", Font.BOLD, 14)
tempname = WindowManager.getUniqueName(self.__img.getShortTitle())
for i in range(len(self.__measures)) :
#print "i", i, self.__measures[i]
yarray=[]
flag=True
miny=10000000000
maxy=-1000000000
#we find the min and max values in order to set the scale.
for cellname in self.__listcellname :
colors.append(dico[cellname].getColor())
yarray.append(self.__dictMeasures[dico[cellname]][self.__measures[i]])
#for meas in self.__dictMeasures[dico[cellname]][self.__measures[i]] :
for meas in yarray[-1] :
if (meas<miny) and (Double.isNaN(meas)==False) :
miny=meas
if max(yarray[-1])>maxy : maxy=max(yarray[-1])
miny-=0.1*miny
maxy+=0.1*maxy
count=0.05
for j in range(len(yarray)) :
if j==0 :
if len(self.__measures)>1 :
plot=Plot("Plots-"+str(self.__measures[i]),namex,str(self.__measures[i]),x,yarray[j])
else :
plot=Plot("Plot-"+tempname,namex,str(self.__measures[i]),x,yarray[j])
plot.setLimits(minx,maxx,miny,maxy)
plot.setColor(colors[j])
plot.changeFont(font)
plot.addLabel(0.05, count, self.__listcellname[j])
else :
plot.setColor(colors[j])
plot.setLineWidth(3)
plot.addPoints(x,yarray[j],Plot.LINE)
plot.addLabel(0.05, count, self.__listcellname[j])
count+=0.05
plot.setColor(colors[0])
plot.show()
if len(self.__measures)>1 :
IJ.run("Images to Stack", "name="+tempname+"-plots title=Plots- use")
#def histbuttonPressed(event) :
#
# pass
# Represents the values in a tab.
def tabbuttonPressed(event) :
tab="\t"
headings=[]
measures=[]
#img=IJ.getImage()
#for i in range(self.__img.getImageStackSize()+1) :
for i in range(self.__maxLife+1) :
headings.append("Slice "+str(i))
headings[0]=WindowManager.getUniqueName(self.__img.getShortTitle())
#for m in self.__measurescompl :
for m in self.__dictMeasures[dico[self.__listcellname[0]]].keys() :
headstring=""
for head in headings:
headstring+=head+tab
tw=TextWindow(self.__listfiles[0]+"-"+m,headstring,"",800,600)
tp=tw.getTextPanel()
#for cellname in dico.keys() :
for cellname in self.__listcellname :
line=[]
line=[str(meas)+tab for meas in self.__dictMeasures[dico[cellname]][m]]
line.insert(0, cellname+tab)
linestr=""
for s in line: linestr+=s
tp.appendLine(linestr)
tp.updateDisplay()
if self.__measuresparambool_global[0] :
tw=TextWindow("Latency","cell\tLatency", "",800,600)
tp=tw.getTextPanel()
for i in range(len(self.__listcellname)) :
#if latencies[i][0] : line=self.__listcellname[i]+"\t"+str(latencies[i][1])
#else : line=self.__listcellname[i]+"\t"+"NaN"
line=self.__listcellname[i]+"\t"+str(latencies[i][1])
tp.appendLine(line)
tp.updateDisplay()
def helpbuttonPressed(event) :
IJ.showMessage("TO DO")
def newsetPressed(event) :
gd0.dispose()
self.__settings()
def alignbuttonPressed(event) :
IJ.showMessage("TO DO")
def mergebuttonPressed(event) :
IJ.showMessage("TO DO")
def saveResults() :
#if len(self.__listcellname) == 0 :
nbslices=self.__maxLife
acell=dico.values()[0]
if self.__useTime :
x = acell.getListTimes()
namex="Time_sec"
else :
x = range(1,nbslices+1)
namex = "Frame"
if not path.exists(self.__rootpath+"Results"+os.path.sep) : os.makedirs(self.__rootpath+os.path.sep+"Results"+os.path.sep, mode=0777)
tab="\t"
nl="\n"
measures=[]
headstring=""
#if self.__savemode : mode = "a"
#else : mode ="w"
mode = "a"
#for i in range(1, self.__maxLife+1) :headstring += "Slice_"+str(i)+tab
for i in range(self.__maxLife) :headstring += str(x[i])+tab
#for m in self.__measurescompl :
for m in self.__dictMeasures[dico[self.__listcellname[0]]].keys() :
f = open(self.__rootpath+"Results"+os.path.sep+m+".txt", mode)
#f.write(m+nl)
f.write(imgName+"-"+self.__time+"-"+m+"-"+namex+tab+headstring+nl)
if len(self.__listcellname) == 0 : f.write("no cells")
else :
for cellname in self.__listcellname :
linestr=cellname+tab
for measure in self.__dictMeasures[dico[cellname]][m] :
#print m, cellname, measure
linestr += str(measure)+tab
linestr+=nl
f.write(linestr)
f.close()
if self.__measuresparambool_global[0] :
m = "Latency"
f = open(self.__rootpath+"Results"+os.path.sep+m+".txt", mode)
f.write(imgName+"-"+self.__time+"-"+m+nl)
for i in range(len(self.__listcellname)) :
#if latencies[i][0] : line=self.__listcellname[i]+"\t"+str(latencies[i][1])
#else : line=self.__listcellname[i]+"\t"+"NaN"
line=self.__listcellname[i]+"\t"+str(latencies[i][1])
line+=nl
f.write(line)
f.close()
#
# ----------- main measures dialog -------------------------
#
# Allows the user to choose the measures to make, etc..
measureslabels_indep=["MaxFeret","MinFeret","AngleFeret","XFeret","YFeret","Area","Angle","Major","Minor","Solidity","AR","Round","Circ","XC","YC","FerCoord","FerAxis","MidAxis"]
measureslabels_dep=["Mean","StdDev","IntDen","Kurt","Skew","XM","YM","Fprofil","MidProfil","NFoci","ListFoci","ListAreaFoci","ListPeaksFoci","ListMeanFoci"]
measureslabels_global=["Latency", "velocity", "cumulatedDist"]
measureslabels_dep_tabonly=set(["MidAxis","FerCoord","FerAxis","Fprofil","MidProfil","ListFoci","ListAreaFoci","ListPeaksFoci","ListMeanFoci"])
ens_measures_global=set(measureslabels_global)
ens_measures_indep=set(measureslabels_indep)
ens_measures_dep=set(measureslabels_dep)
measureslabels=[]
for label in measureslabels_indep :
measureslabels.append(label)
for label in measureslabels_dep :
measureslabels.append(label)
#self.__defaultmeasures=[False for i in range(len(measureslabels))]
#self.__defaultmeasures_global=[False for i in range(len(measureslabels_global))]
gdmeasures=NonBlockingGenericDialog("MeasuresChoice")
gdmeasures.setFont(Font("Courrier", 1, 10))
gdmeasures.addMessage("******* TIME SETTINGS *******")
gdmeasures.addCheckbox("Only starting at begining :", self.__onlystart) # 1 only start
gdmeasures.addNumericField("Minimal Lifetime : ",self.__minLife,0)
gdmeasures.addNumericField("Maximal Lifetime : ",self.__maxLife,0)
#gdmeasures.addNumericField("Maximal Lifetime : ",self.__img.getImageStackSize(),0)
gdmeasures.addCheckbox("x axis in seconds", self.__useTime) # 2 use time
gdmeasures.addMessage("")
gdmeasures.addMessage("")
gdmeasures.addMessage("Choose the measures to make on the cells : ")
gdmeasures.addMessage("******* TIME MEASURES *******")
gdmeasures.addCheckboxGroup(4,8,measureslabels,self.__defaultmeasures)
gdmeasures.addMessage("")
gdmeasures.addMessage("******* GLOBAL MEASURES *******")
gdmeasures.addMessage("PLEASE : If you have selected movement parameters you MUST to select XC and YC !")
gdmeasures.addCheckboxGroup(3,1,measureslabels_global,self.__defaultmeasures_global)
gdmeasures.addNumericField("Noise value for maxima finder: ",self.__noise,0)
gdmeasures.addMessage("")
gdmeasures.addMessage("******* OPTIONS *******")
gdmeasures.addCheckbox("Select the cells in next dialog ?", self.__onlyselect) # 3 only select
gdmeasures.addCheckbox("Save results to text files ?", self.__savetables) # 4 save files
#gdmeasures.addCheckbox("Append mode ?", self.__savemode) # 5 append mode
gdmeasures.addCheckbox("Analyse in batch mode ?", self.__batchanalyse) # 6 analysis batch mode
gdmeasures.addCheckbox("Update overlay ?", self.__updateoverlay) # 7 update overlay
gdmeasures.addMessage("")
gdmeasures.addMessage("")
help_panel=Panel()
helpbutton=Button("HELP")
helpbutton.actionPerformed = helpbuttonPressed
help_panel.add(helpbutton)
gdmeasures.addPanel(help_panel)
gdmeasures.hideCancelButton()
if not self.__batchanalyse :
gdmeasures.showDialog()
self.__onlystart=gdmeasures.getNextBoolean() # 1 only start
self.__minLife=gdmeasures.getNextNumber()
self.__maxLife=gdmeasures.getNextNumber()
self.__useTime=gdmeasures.getNextBoolean() # 2 use time
self.__measuresparambool=[]
self.__measuresparambool_global=[]
for i in range(len(measureslabels)) :
self.__measuresparambool.append(gdmeasures.getNextBoolean())
self.__defaultmeasures[i]=self.__measuresparambool[-1]
for i in range(len(measureslabels_global)) :
self.__measuresparambool_global.append(gdmeasures.getNextBoolean())
self.__defaultmeasures_global[i] = self.__measuresparambool_global[i]
self.__noise=gdmeasures.getNextNumber()
self.__onlyselect=gdmeasures.getNextBoolean() # 3 only select
self.__savetables = gdmeasures.getNextBoolean() # 4 save files
#self.__savemode = gdmeasures.getNextBoolean() # 5 append mode
self.__batchanalyse = gdmeasures.getNextBoolean() # 6 analyse mode
self.__updateoverlay = gdmeasures.getNextBoolean() # 7 update overlay
# we update a list of all cells that have a lifetime corresponding to what the user chose.
if len (self.__allcells) == 0 :
for cellname in dico.keys() :
if dico[cellname].getLifeTime()>=self.__minLife : #and dico[cellname].getLifeTime()<=self.__maxLife :
if self.__onlystart :
if dico[cellname].getSlideInit()<2 : self.__allcells.append(cellname)
else : self.__allcells.append(cellname)
if self.__noise == 0 : self.__noise = None
if self.__batchanalyse : self.__onlyselect = False
if self.__onlyselect :
try :
self.__gw
except AttributeError :
if not path.exists(self.__pathdir+"Selected-Cells"+os.path.sep) : os.makedirs(self.__pathdir+os.path.sep+"Selected-Cells"+os.path.sep, mode=0777)
self.__gw = CellsSelection()
self.__gw.setTitle(imgName)
self.__gw.run(self.__allcells, self.__pathdir+"ROIs"+os.path.sep)
self.__gw.show()
self.__gw.setSelected(self.__allcells)
while not self.__gw.oked and self.__gw.isShowing() :
self.__gw.setLabel("Validate selection with OK !!")
self.__listcellname = list(self.__gw.getSelected())
self.__gw.resetok()
self.__gw.setLabel("...")
self.__gw.hide()
else :
if self.__gw.getTitle() == imgName :
self.__gw.show()
self.__gw.setSelected(self.__listcellname)
self.__listcellname[:]=[]
while not self.__gw.oked and self.__gw.isShowing() :
self.__gw.setLabel("Validate selection with OK !!")
self.__listcellname = list(self.__gw.getSelected())
self.__gw.resetok()
self.__gw.setLabel("...")
self.__gw.hide()
else :
self.__gw.dispose()
if not path.exists(self.__pathdir+"Selected-Cells"+os.path.sep) : os.makedirs(self.__pathdir+os.path.sep+"Selected-Cells"+os.path.sep, mode=0777)
self.__gw = CellsSelection()
self.__gw.setTitle(imgName)
self.__gw.run(self.__allcells, self.__pathdir+"ROIs"+os.path.sep)
self.__gw.show()
self.__gw.setSelected(self.__allcells)
self.__listcellname[:]=[]
while not self.__gw.oked and self.__gw.isShowing() :
self.__gw.setLabel("Validate selection with OK !!")
self.__listcellname = list(self.__gw.getSelected())
self.__gw.resetok()
self.__gw.setLabel("...")
self.__gw.hide()
filestodelet=glob.glob(self.__pathdir+"Selected-Cells"+os.path.sep+"*.cell")
for f in filestodelet : os.remove(f)
for cell in self.__listcellname :
sourcestr = self.__pathdir+"Cells"+os.path.sep+cell+".cell"
deststr = self.__pathdir+"Selected-Cells"+os.path.sep+cell+".cell"
#os.system("copy "+sourcestr+", "+deststr)
#shutil.copy(self.__pathdir+"Cells"+os.path.sep+cell+".cell",self.__pathdir+"Selected-Cells"+os.path.sep+cell+".cell")
shutil.copy(sourcestr,deststr)
self.__dictNcells[imgName] = len(self.__listcellname)
else :
self.__listcellname = list(self.__allcells)
self.__dictNcells[imgName] = len(self.__listcellname)
if len(self.__listcellname) == 0 :
self.__dictNcells[imgName] = 0
return False
self.__img.hide()
# we make the measures.
for i in range(len(measureslabels)) :
IJ.showProgress(i, len(measureslabels))
if self.__measuresparambool[i]==True :
self.__measurescompl.append(measureslabels[i])
if (measureslabels[i] in measureslabels_dep_tabonly)==False :
self.__measures.append(measureslabels[i])
if (i<18) and (measureslabels[i] in ens_measures_indep) :
self.__measureAll(self.__img,measureslabels[i],False, imgName, self.__noise)
ens_measures_indep.discard(measureslabels[i])
if i>=18 :
self.__measureAll(self.__img,measureslabels[i],True, imgName, self.__noise)
if self.__measuresparambool_global[0] : # calculate latency
latencies=[]
for i in range(len(self.__listcellname)) :
IJ.showProgress(i, len(self.__listcellname))
latencies.append(self.latencie(self.__listcellname[i], self.__img, imgName, self.__useTime))
if self.__measuresparambool_global[1] : # calculate velocity
self.__measures.append("velocity")
#velocities=[]
for i in range(len(self.__listcellname)) :
IJ.showProgress(i, len(self.__listcellname))
self.__measureVelocity(self.__img,imgName)
if self.__measuresparambool_global[2] : # calculate cumulatedDistance
self.__measures.append("cumulatedDist")
#velocities=[]
for i in range(len(self.__listcellname)) :
IJ.showProgress(i, len(self.__listcellname))
self.__measurecumulDist(self.__img,imgName)
self.__img.show()
self.__img.getProcessor().resetThreshold()
if self.__updateoverlay :
if self.__img.getOverlay() is not None : self.__img.getOverlay().clear()
outputrois=[]
cellnames=[]
self.__img.hide()
for cellname in self.__listcellname :
for r in dico[cellname].getListRoi():
if isinstance(r,Roi) :
pos=r.getPosition()
#print "MC overlay", cellname, r.getName(), pos
#r.setPosition(0)
#overlay.add(r)
outputrois.append(r)
if "cell" in r.getName() : cellnames.append(r.getName())
else : cellnames.append(str(pos)+"-"+cellname)
#print cellnames[-1]
rm = RoiManager.getInstance()
if (rm==None): rm = RoiManager()
rm.show()
self.__img.show()
IJ.selectWindow(self.__img.getTitle())
rm.runCommand("reset")
for i in range(len(outputrois)) :
outputrois[i].setName(cellnames[i])
rm.addRoi(outputrois[i])
rm.select(rm.getCount()-1)
rm.runCommand("Rename", cellnames[i])
IJ.run("Show Overlay", "")
rm.runCommand("UseNames", "true")
rm.runCommand("Associate", "true")
IJ.run(self.__img, "Labels...", "color=red font=12 show use")
IJ.run(self.__img, "From ROI Manager", "")
rm.runCommand("Show None")
rm.runCommand("Show All")
# ----------- batch analyse ------------------------
if self.__batchanalyse :
if self.__savetables : saveResults()
self.__dictMeasures.clear()
self.__allcells[:]=[]
self.__measurescompl[:]=[]
self.__measures[:]=[]
return False
# ---------- display methodes dialog ----------------
# Allows the user to choose how to see the results of the measures.
gd0=NonBlockingGenericDialog("Display")
gd0.addMessage("How do you want to see the results ?")
panel0=Panel()
diagrambutton=Button("Diagram")
diagrambutton.actionPerformed = diagrambuttonPressed
panel0.add(diagrambutton)
graphbutton=Button("Graph")
graphbutton.actionPerformed = graphbuttonPressed
panel0.add(graphbutton)
tabbutton=Button("Tab")
tabbutton.actionPerformed = tabbuttonPressed
panel0.add(tabbutton)
gd0.addPanel(panel0)
gd0.addCheckbox("Analyse next stack ?", self.__nextstack)
gd0.hideCancelButton()
gd0.showDialog()
self.__nextstack = gd0.getNextBoolean()
# ---------- save tables ---------------------------
if self.__savetables : saveResults()
# --------- re-start analysis -------------------
self.__dictMeasures.clear()
#self.__listcellname[:]=[]
self.__allcells[:]=[]
self.__measurescompl[:]=[]
self.__measures[:]=[]
if self.__nextstack : return False
else : return True
def __ImportCells(self, imagesnames) :
#self.__dictCells[imgName]={}
rm = RoiManager.getInstance()
if (rm==None): rm = RoiManager()
rm.runCommand("reset")
listpaths = []
listfilescells=[]
if self.__optionImages :
IJ.showMessage("Select the folder 'Cells' containing the cells to import")
selectdir=IJ.getDirectory("image")
selectdir=IJ.getDirectory("")
listfilescells.extend(glob.glob(selectdir+os.path.sep+"*"))
listpaths.append("")
else :
IJ.showMessage("Select the text file containing the list cell paths (listpaths.txt)")
selectdir=IJ.getDirectory("current")
frame = Frame("Text file settings ?")
fd = FileDialog(frame)
fd.setDirectory(selectdir)
fd.show()
selectdir = fd.getDirectory()
textfile = fd.getFile()
fichier = open(selectdir+textfile,"r")
listpaths=[ glob.glob(f.split("\n")[0]+"Selected-Cells"+os.path.sep+"*") for f in fichier.readlines()]
#for f in templist :
# listpaths.append(f.split("\n")+"Cells")
listfilescells.append("")
if listfilescells[0]=="" : importmode = True
else : importmode = False
for j in range(len(listpaths)) :
self.__dictCells[imagesnames[j]]={}
if importmode : listfilescells = listpaths[j]
pathtemp = []
for cellfile in listfilescells :
filetemp = open(cellfile,"r")
linestemp=filetemp.readlines()
for line in linestemp :
params=line.split("=")
values=params[1].split("\n")
if params[0] == "NAMECELL" :
celltemp=Bacteria_Cell(str(values[0]))
self.__dictCells[imagesnames[j]][values[0]]=celltemp
self.__dictMeasures[self.__dictCells[imagesnames[j]][values[0]]]={}
if params[0] == "PATHROIS" :
pathtemp.append(str(values[0]))
if params[0] == "NSLICES" :
for i in range(int(values[0])) :
celltemp.getListRoi().append("")
if params[0] == "SLICEINIT" :
celltemp.setSlideInit(int(values[0]))
for i in range(int(values[0])-2) :
celltemp.setRoi("NOT HERE YET",i)
if params[0] == "SLICEEND" :
celltemp.setSlideEnd(int(values[0]))
for i in range(int(values[0])) :
celltemp.setRoi("LOST",i)
if params[0] == "COLOR" :
colorstemp=values[0].split(";")
celltemp.setColor(Color(int(colorstemp[0]),int(colorstemp[1]),int(colorstemp[2])))
indiceroi=0
ind=0
tempimp = WindowManager.getImage(imagesnames[j])
if tempimp is not None :
IJ.selectWindow(imagesnames[j])
tempimp.show()
else :
if imagesnames[j][-4:]==".tif" :
IJ.selectWindow(imagesnames[j][:-4])
tempimp = IJ.getImage()
else :
IJ.selectWindow(imagesnames[j]+".tif")
tempimp = IJ.getImage()
rm.runCommand("reset")
for cellname in self.__dictCells[imagesnames[j]].keys() :
rm.runCommand("Open", pathtemp[ind])
ind+=1
nbtemp=self.__dictCells[imagesnames[j]][cellname].getLifeTime()
for i in range(nbtemp) :
rm.select(tempimp, indiceroi)
roi=rm.getSelectedRoisAsArray()[0]
self.__dictCells[imagesnames[j]][cellname].setRoi(roi,i+self.__dictCells[imagesnames[j]][cellname].getSlideInit()-1)
indiceroi+=1
IJ.run("Show Overlay", "")
rm.runCommand("UseNames", "true")
rm.runCommand("Associate", "true")
IJ.run(tempimp, "Labels...", "color=red font=12 show use")
if rm.getCount()>0 : IJ.run(tempimp, "From ROI Manager", "")
rm.runCommand("Show None")
rm.runCommand("Show All")
roipath = os.path.split(pathtemp[0])[0]+os.path.sep
rootpath = roipath.rsplit(os.path.sep, 2)[0]+os.path.sep
self.__listpaths[j] = rootpath
self.__rootpath=rootpath
def adjustRoiAndMeasure(imp, frameNumber, dstDir):
rm = RoiManager.getInstance()
if not rm:
rm = RoiManager()
nROIs = rm.getCount()
indexlist = range(nROIs)
if nROIs > 2:
for roi in indexlist:
indexlist_copy = list(indexlist)
del indexlist_copy[roi]
rm.setSelectedIndexes(indexlist_copy)
rm.runCommand(imp, "Combine")
IJ.run(imp, "Make Inverse", "")
rm.addRoi(imp.getRoi())
new_nROI = rm.getCount()
rm.setSelectedIndexes([roi, new_nROI - 1])
rm.runCommand(imp, "AND")
if imp.getRoi():
rm.addRoi(imp.getRoi())
rm.setSelectedIndexes([new_nROI])
rm.runCommand("Rename", "Cell" + str(roi))
else:
rm.setSelectedIndexes([roi])
rm.addRoi(imp.getRoi())
rm.setSelectedIndexes([new_nROI - 1])
rm.runCommand(imp, "Delete")
elif nROIs == 2:
for roi in indexlist:
indexlist_copy = list(indexlist)
del indexlist_copy[roi]
rm.setSelectedIndexes(indexlist_copy)
IJ.run(imp, "Make Inverse", "")
rm.addRoi(imp.getRoi())
new_nROI = rm.getCount()
rm.setSelectedIndexes([roi, new_nROI - 1])
rm.runCommand(imp, "AND")
rm.addRoi(imp.getRoi())
rm.setSelectedIndexes([new_nROI])
rm.runCommand("Rename", "Cell" + str(roi))
rm.setSelectedIndexes([new_nROI - 1])
rm.runCommand(imp, "Delete")
elif nROIs == 1:
nROIs = 0
new_nROI = 1
rm.setSelectedIndexes([new_nROI])
rm.runCommand("Rename", "Cell0")
else:
return
adjustedROIs = range(nROIs, new_nROI, 1)
rm.setSelectedIndexes(adjustedROIs)
measureChannels(adjustedROIs, imp, frameNumber)
rm.setSelectedIndexes(adjustedROIs)
rm.runCommand("Save selected",
dstDir + "\\Frame " + str(frameNumber + 1) + " roi set.zip")
rm.runCommand(imp, "Deselect")
rm.runCommand(imp, "Delete")
num_frames = imp.getNFrames();
num_z_stacks = imp.getNSlices();
if num_frames < num_z_stacks:
imp.setDimensions(1,num_frames,num_z_stacks);
ROI_list = os.listdir(Folder_ROIs);
Roi_File = Folder_ROIs + ROI_list[ROI_idx];
print(ROI_list[ROI_idx]);
rm = RoiManager();
rm = RoiManager.getInstance();
rm.runCommand("Open", Roi_File);
total_rois = rm.getCount();
for idx in range(total_rois):
#----------------------------
# Create the model object now
#----------------------------
# Some of the parameters we configure below need to have
# a reference to the model at creation. So we create an
# empty model now.
model = Model();
# Send all messages to ImageJ log window.
model.setLogger(Logger.IJ_LOGGER);
model.setPhysicalUnits(Calib.getUnit(), Calib.getTimeUnit());
def merge_incorrect_splits_and_get_centroids(imp,
centroid_distance_limit=100,
size_limit=100):
"""if particles are found with centroids closer than centroid_distance_limit and both have size<size_limit, get average centroid"""
imp.killRoi()
rt = ResultsTable()
out_imp = IJ.createImage("Nuclei_centroids_from_{}".format(imp.getTitle()),
imp.getWidth(), imp.getHeight(), 1, 8)
IJ.run(out_imp, "Select All", "")
IJ.run(out_imp, "Set...", "value=0 slice")
cal = imp.getCalibration()
mxsz = imp.width * cal.pixelWidth * imp.height * cal.pixelHeight
roim = RoiManager(False)
pa = ParticleAnalyzer(
ParticleAnalyzer.ADD_TO_MANAGER, ParticleAnalyzer.AREA
| ParticleAnalyzer.SLICE | ParticleAnalyzer.CENTROID, rt, 0,
size_limit)
pa.setRoiManager(roim)
roim.reset()
rt.reset()
pa.analyze(imp)
centroids_set = set()
if roim.getCount() > 0:
rt_xs = rt.getColumn(rt.getColumnIndex("X")).tolist()
rt_ys = rt.getColumn(rt.getColumnIndex("Y")).tolist()
centroids = [(x, y) for x, y in zip(rt_xs, rt_ys)]
for c in centroids:
ds = [
math.sqrt((c[0] - cx)**2 + (c[1] - cy)**2)
for (cx, cy) in centroids
]
close_mask = [d < centroid_distance_limit for d in ds]
# if no other centroids are within centroid_distance_limit, add this centroid to the output set
# otherwise, add the average position of this centroid and those within centroid_distance_limit to the output set
centroids_set.add(
(sum([msk * b[0] for msk, b in zip(close_mask, centroids)]) /
sum(close_mask),
sum([msk * b[1] for msk, b in zip(close_mask, centroids)]) /
sum(close_mask)))
roim.reset()
rt.reset()
pa = ParticleAnalyzer(
ParticleAnalyzer.ADD_TO_MANAGER, ParticleAnalyzer.AREA
| ParticleAnalyzer.SLICE | ParticleAnalyzer.CENTROID, rt, size_limit,
mxsz)
pa.setRoiManager(roim)
pa.analyze(imp)
if roim.getCount() > 0:
if rt.columnExists("X"):
rt_xs = rt.getColumn(rt.getColumnIndex("X")).tolist()
rt_ys = rt.getColumn(rt.getColumnIndex("Y")).tolist()
centroids = [(x, y) for x, y in zip(rt_xs, rt_ys)]
for c in centroids:
centroids_set.add(c)
centroids = list(centroids_set)
# cal = imp.getCalibration()
centroids = [(c[0] / cal.pixelWidth, c[1] / cal.pixelHeight)
for c in centroids]
roim.reset()
roim.close()
for idx, c in enumerate(centroids):
roi = OvalRoi(c[0], c[1], 1, 1)
out_imp.setRoi(roi)
IJ.run(out_imp, "Set...", "value={} slice".format(idx + 1))
#imp.changes = False
#imp.close()
return out_imp, centroids
pa = ParticleAnalyzer((ParticleAnalyzer.ADD_TO_MANAGER | ParticleAnalyzer.SHOW_MASKS), (Measurements.CENTROID | Measurements.STACK_POSITION), rt, 500, 30000, 0.0, 1.0)
pa.setHideOutputImage(False)
keep_rois = [];
pa.analyze(imp);
IJ.run("Set Measurements...", "centroid redirect=None decimal=3");
frames = imp.getNFrames();
for fridx in range(0, frames):
rt.reset();
imp.setSliceWithoutUpdate(fridx + 1);
ip = imp.getProcessor();
if not pa.analyze(imp, ip):
raise Exception("something went wrong analysing particles!")
rt.show("centroids");
rm = RoiManager.getInstance();
if rm.getCount() > 0:
rois = rm.getRoisAsArray();
centroidsx = rt.getColumn(rt.getColumnIndex('X'));
centroidsy = rt.getColumn(rt.getColumnIndex('Y'));
print(centroidsx);
print(centroidsy);
gd = GenericDialog("Continue?");
gd.showDialog();
if gd.wasCanceled():
raise Exception("Run interupted");
for roi in rois:
imp.setRoi(roi);
stats = ImageStatistics().getStatistics(ip);
print(stats.xCenterOfMass)
def Overlayer(org_size, dirs):
""" Overlays ROIs with appropriate color,
saves to .tif and animates aligned images to .gif """
# Get colors.
Colors, Colors_old = colorlist()
# Get ROImanager.
rm = RoiManager().getInstance()
rois = rm.getCount()
# Overlays ROI on aligned images, converts to 8-bit (for gif).
for root, directories, filenames in os.walk(dirs["Composites_Aligned"]):
for filename in filenames:
imp = IJ.openImage(os.path.join(root, filename))
converter = ImageConverter(imp)
converter.setDoScaling(True)
converter.convertToGray8()
# Lookup table and local contrast enhancement for vizualisation.
IJ.run(imp, "Rainbow RGB", "")
IJ.run(imp, "Enhance Local Contrast (CLAHE)",
"blocksize=127 histogram=256 maximum=3 mask=*None*")
for roi in range(rois):
roi_obj = rm.getRoi(roi)
roi_obj.setStrokeWidth(2)
if roi < 19:
roi_obj.setStrokeColor(Color(*Colors[roi][0:3]))
else:
roi_obj.setStrokeColor(eval(Colors_old[roi]))
rm.moveRoisToOverlay(imp)
IJ.saveAs(imp, "Tiff", os.path.join(dirs["Overlays"], filename))
# Opens overlaid images, saves as tiff stack.
overlay_stack = IJ.run("Image Sequence...", "open="+dirs["Overlays"]+
" number=3040 starting=0 increment=1 scale=300 file=.tif sort")
# Takes care of spaces in titles.
tiftitle = Title.replace(" ", "_")
tiftitle = tiftitle.replace(".", "_")
# Gets dimensions for scalebar.
imp = WindowManager.getImage("Overlays")
dimensions = imp.getDimensions()
size = dimensions[0] + dimensions[1]
microns = org_size / size
# Sets scale and writes scale-bar, flattens overlays.
IJ.run(imp, "Set Scale...", "distance=1 known="
+str(microns)+" pixel=1 unit=micron")
IJ.run(imp, "Scale Bar...",
"width=10 height=4 font=14 color=Yellow background=None location=[Lower Right] bold overlay")
IJ.run(imp, "Flatten", "stack")
IJ.saveAs(imp, "Tiff", os.path.join(dirs["Gifs"], tiftitle))
# Animates tiff stack from directoy.
for root, directories, filenames in os.walk(dirs["Gifs"]):
for filename in filenames:
if tiftitle in filename and filename.endswith(".tif"):
# set=xx parameter controls gif speed.
# for additional parameters run with macro recorder.
try:
print "Animating gif..."
imp = WindowManager.getImage(tiftitle + ".tif")
gif = IJ.run("Animated Gif ... ",
"set=200 number=0 filename="
+ os.path.join(dirs["Gifs"], tiftitle + ".gif"))
except Exception, e:
print str(e)
print "gif animated."
def tethered_cell(image_path, frame_number=100, frame_rate=100.0, CCW=1):
"""
parameter setting; frame rate (frame/sec)
CCW = 1 : the motor rotation direction and the cell rotation direction on the image are same
CCW = -1: the motor rotation direction and the cell rotation direction on the image are different
"""
opener = Opener()
imp = opener.openImage(image_path)
image_slice_number = imp.getNSlices()
rm = RoiManager().getInstance()
if image_slice_number < frame_number: # too short movie
IJ.log('Number of frame of the movie is fewer than the number of frame that you selected')
return False
# create result directory
result_path = image_path + '_tethered_cell_result'
if os.path.lexists(result_path) is False:
os.mkdir(result_path)
#z projection; standard deviation, tethered cell shorws circle
IJ.run(imp, 'Subtract Background...', 'rolling=5 light stack')
IJ.run(imp, 'Median...', 'radius=2 stack')
IJ.run(imp, 'Z Project...', 'stop=500 projection=[Standard Deviation]')
zimp = IJ.getImage()
IJ.saveAs(zimp, 'bmp', os.path.join(result_path,'STD_DEV.bmp'))
# pick up tethered cell
IJ.setAutoThreshold(zimp, 'MaxEntropy dark')
IJ.run(zimp, 'Convert to Mask', '')
IJ.run('Set Measurements...', "area centroid bounding shape feret's limit redirect=None decimal=3")
IJ.run(zimp, 'Analyze Particles...', 'size=30-Infinity circularity=0.88-1.00 show=Nothing display exclude clear include')
zrt = ResultsTable.getResultsTable()
IJ.saveAs('Results', os.path.join(result_path,'RoiInfo.csv'))
#tcX and tcY are xy coordinates of tethered cell, tcdia is outer diameter of rotating tethered cell
#add ROI into stack image
for i in range(zrt.getCounter()):
tcX = zrt.getValue('X', i)
tcY = zrt.getValue('Y', i)
tcdia = zrt.getValue('Feret', i)
rm.add(imp, OvalRoi(tcX - tcdia/2.0, tcY - tcdia/2.0, tcdia + 1, tcdia + 1), i)
#calculate rotation speed by ellipse fitting
IJ.setAutoThreshold(imp, 'Li')
for roi_number in range(rm.getCount()):
t = []
XM = []
YM = []
theta = []
rotation_speed = []
area = []
imp.setRoi(rm.getRoi(roi_number))
cropped_imp = Duplicator().run(imp)
IJ.run('Set Measurements...', 'area mean center fit limit redirect=None decimal=3')
rm.select(roi_number)
rt = rm.multiMeasure(imp)
# check cell is present while analysis. Don't a cell gose anywhare?
for i in range(frame_number):
area.append(rt.getValue('Area1', i))
if 0 in area:
continue
for i in range(frame_number):
t.append((1/frame_rate)*i)
XM.append(rt.getValue('XM1', i))
YM.append(rt.getValue('YM1', i))
theta.append(rt.getValue('Angle1', i)/180.0*math.pi) # convert to radian
if i == 0:
rotation_speed.append(0)
else:
# phase treatment, theta should be -pi ~ pi
temp_rotation_speed = [theta[i] - theta[i-1],
theta[i] - theta[i-1] + math.pi,
theta[i] - theta[i-1] - math.pi,
theta[i] - theta[i-1] + 2*math.pi,
theta[i] - theta[i-1] - 2*math.pi]
temp_rotation_speed = sorted(temp_rotation_speed, key = lambda x :abs(x) )[0]
rotation_speed.append(CCW*temp_rotation_speed/(2.0*math.pi)*frame_rate)
# write csv
# earch columns indicate 1:index, 2:time(sec), 3:X-coordinate of center of mass(pixel), 4:Y-coordinate of center of mass (pixel), 5:Angle(Radian), 6:Rotation Speed(Hz)
with open(os.path.join(result_path,'Roi' + str(roi_number) + '.csv'), 'w') as f:
writer = csv.writer(f)
writer.writerow(['Index', 'time(s)', 'X', 'Y', 'Angle(rad)', 'Rotation Speed(Hz)'])
for i in range(len(t)):
writer.writerow([i, t[i], XM[i], YM[i], theta[i], rotation_speed[i]])
# plot x-y, t-x, t-y, t-rotation speed, save plot as bmp
plotRotation(roi_number, result_path, t, XM, YM, rotation_speed)
IJ.saveAs(cropped_imp, 'tiff', os.path.join(result_path,'Roi' + str(roi_number) + '.tiff'))
rt.reset()
# get analysis date and time
dt = datetime.datetime.today()
dtstr = dt.strftime('%Y-%m-%d %H:%M:%S')
# wtite analysis setting
with open(os.path.join(result_path,'analysis_setting.csv'), 'w') as f:
writer = csv.writer(f)
writer.writerow(['Analysis Date','frame number','frame rate','CCW direction', 'Method','Auto threshold', 'Subtruct Background', 'Median filter'])
writer.writerow([dtstr, frame_number, frame_rate, CCW, 'Ellipse', 'Li', '5.0', '2'])
# save roi
if rm.getCount() != 0:
rm.runCommand('Save', os.path.join(result_path, 'Roi.zip'))
zimp.close()
imp.close()
rm.close()
zrt.reset()
IJ.run(
"Analyze Particles...",
"size=" + str(uGsize) + "-Infinity pixel circularity=" +
str(uGshape) + "-" + str(uGshapemax) +
" show=[Overlay Masks] exclude clear summarize add")
print "ATP, Exclude edges"
else:
IJ.run(
"Analyze Particles...", "size=" + str(uGsize) +
"-Infinity pixel circularity=" + str(uGshape) + "-" +
str(uGshapemax) + " show=[Overlay Masks] clear summarize add")
print "ATP, include all"
# ROI management
rm.runCommand("Combine")
rm.runCommand("Add")
rct = rm.getCount()
fullTitle = ImagePlus.getTitle(thisImage)
shortTitle = ImagePlus.getShortTitle(thisImage)
IJ.run("Invert LUT") ### Invert LUT
rm.rename(rct - 1, str(shortTitle))
rm.select(rct - 1)
if image == newPaths[0]:
rm.runCommand(
"Save",
analysisOut + File.separatorChar + "ROIindividual_wt" + ".zip")
if image == newPaths[1]:
rm.runCommand(
"Save",
analysisOut + File.separatorChar + "ROIindividual_dic" + ".zip")
if image == newPaths[2]:
rm.runCommand(