def drawLines(imp, points=None):
if points and (len(points) % 2 == 0):
# points is numeric list of even length
pRoi = PointRoi(points[0::2], points[1::2], len(points) / 2)
pRoi.setShowLabels(True)
pRoi.setSize(3)
imp.setRoi(pRoi)
roi = imp.getRoi()
pp = roi.getFloatPolygon()
# print "Added", pp.npoints
if pp.npoints <= 1:
# don't draw if only one point
return
xys = []
for i in xrange(pp.npoints):
xys.append([pp.xpoints[i], pp.ypoints[i]])
ol = Overlay()
x0 = xys[0][0]
y0 = xys[0][1]
cal = imp.getCalibration()
for i in xrange(1, pp.npoints):
xi = xys[i][0]
yi = xys[i][1]
# prepare text label
d = math.sqrt((xi - x0)**2 + (yi - y0)**2) * cal.pixelWidth
dText = String.format("%.2f ", d) + cal.getUnits()
textOffset = 30
xt = xi
yt = yi
# if xi > x0:
# xt += textOffset
if xi < x0:
xt -= textOffset
# if yi > y0:
# yt += textOffset
if yi < y0:
yt -= textOffset
dTextRoi = TextRoi(xt, yt, dText)
ol.add(dTextRoi)
lineRoi = Line(x0, y0, xi, yi)
lineRoi.setStrokeWidth(1)
lineRoi.setStrokeColor(Color(255, 255, 0))
ol.add(lineRoi)
imp.setOverlay(ol)
imp.updateAndDraw()
def drawLines(imp, points=None):
if points and (len(points)%2 == 0):
# points is numeric list of even length
pRoi = PointRoi(points[0::2], points[1::2], len(points)/2)
pRoi.setShowLabels(True)
pRoi.setSize(3)
imp.setRoi(pRoi)
roi = imp.getRoi()
pp = roi.getFloatPolygon()
# print "Added", pp.npoints
if pp.npoints <= 1:
# don't draw if only one point
return
xys = []
for i in xrange(pp.npoints):
xys.append([pp.xpoints[i], pp.ypoints[i]])
ol = Overlay()
x0 = xys[0][0]
y0 = xys[0][1]
cal = imp.getCalibration()
for i in xrange(1, pp.npoints):
xi = xys[i][0]
yi = xys[i][1]
# prepare text label
d = math.sqrt((xi - x0)**2 + (yi - y0)**2) * cal.pixelWidth
dText = String.format("%.2f ", d) + cal.getUnits()
textOffset = 30
xt = xi
yt = yi
# if xi > x0:
# xt += textOffset
if xi < x0:
xt -= textOffset
# if yi > y0:
# yt += textOffset
if yi < y0:
yt -= textOffset
dTextRoi = TextRoi(xt, yt, dText)
ol.add(dTextRoi)
lineRoi = Line(x0, y0, xi, yi)
lineRoi.setStrokeWidth(1)
lineRoi.setStrokeColor(Color(255,255,0))
ol.add(lineRoi)
imp.setOverlay(ol)
imp.updateAndDraw()
def getCells(dicStack):
outStack = ImageStack(W,H)
cells = [None for t in range(T+1)]
for t in range(1,T+1):
mapp = dicStack.getProcessor(t).convertToFloatProcessor()
mapp.subtract( mapp.getStatistics().mean )
mapp.abs()
RankFilters().rank(mapp, 1.0, RankFilters.VARIANCE)
mapp.sqrt()
mapp.blurGaussian(5)
hist = mapp.getHistogram(256)
stats = mapp.getStatistics()
thresh = AutoThresholder().getThreshold( AutoThresholder.Method.Otsu, hist )
thresh = (thresh/float(255)) * (stats.max-stats.min) + stats.min
mask = ByteProcessor(W,H)
for i in range(W*H):
value = mapp.getf(i)
bite = 255 if value>=thresh else 0
mask.set(i, bite)
fillHoles(mask)
ed = 3
for e in range(ed): mask.erode(1, 0)
for d in range(ed): mask.dilate(1, 0)
watershed(mask)
minA = 5000 #px²
mask.setThreshold(255,255, ImageProcessor.NO_LUT_UPDATE)
composite = ThresholdToSelection().convert(mask)
rois = ShapeRoi(composite).getRois()
keep = []
for roi in rois:
if roi.getStatistics().area >= minA:
if not onEdge(roi):
keep.append(roi)
else:
edgeRoi = ShapeRoi(roi)
edgeRoi.setPosition(0,0,t)
edgeRoi.setStrokeColor(Color.YELLOW)
ol.add(edgeRoi)
print("T"+str(t)+" using "+str(len(keep))+"/"+str(len(rois))+" ROIs")
rois = keep
#rois = [ roi for roi in rois if roi.getStatistics().area >= minA and not onEdge(roi) ] #keep big enough and not on edges
# if there is only one Roi, cut it along the fitted ellipse minor axis
if len(rois)==1:
el = EllipseFitter()
mask.setRoi(rois[0])
el.fit(mask, None)
el.makeRoi(mask)
theta = el.angle * (maths.pi/180.0)
length = el.major/2.0
dy = maths.sin(theta)* length
dx = maths.cos(theta)* length
#major axis
lineX0 = el.xCenter - dx
lineY0 = el.yCenter + dy
lineX1 = el.xCenter + dx
lineY1 = el.yCenter - dy
line = Line(lineX0, lineY0, lineX1, lineY1)
line.setStrokeColor(Color.BLUE)
line.setStrokeWidth(1)
line.setPosition(0,0,t)
ol.add(line)
#minor axis scaled length to make sure cut ends are outside Roi
cutX0 = el.xCenter + dy*100
cutY0 = el.xCenter + dx*100
cutX1 = el.yCenter - dy*100
cutY1 = el.yCenter - dx*100
cut = Line(cutX0,cutY0, cutX1, cutY1)
cut.setStrokeWidth(2)
cut = PolygonRoi( cut.getFloatPolygon(), PolygonRoi.POLYGON )
mask.setColor(0)
mask.fill(cut)
composite = ThresholdToSelection().convert(mask)
rois = ShapeRoi(composite).getRois()
rois = [ roi for roi in rois if roi.getStatistics().area >= minA ]
print(str(t) + ":" + str(len(rois)))
rois = [ PolygonRoi(roi.getInterpolatedPolygon(20, True), PolygonRoi.POLYGON) for roi in rois ]
rois = [ PolygonRoi(roi.getConvexHull(), PolygonRoi.POLYGON) for roi in rois ]
rois = sorted(list(rois), key=lambda roi:roi.getLength() ) #size order
rois = rois[-2:] #keep 2 biggest
rois = sorted(list(rois), key=lambda roi:roi.getStatistics().xCentroid+roi.getStatistics().yCentroid ) #top left to bottom right order
if len(rois)>0:
rois[0].setStrokeColor(Color.RED)
rois[0].setPosition(0, 0, t)
ol.add(rois[0])
if len(rois)>1:
rois[1].setStrokeColor(Color.GREEN)
rois[1].setPosition(0, 0, t)
ol.add(rois[1])
cells[t] = (rois[0], rois[1])
return cells
def __midline(self):
debug=False
#print "line 251", self.__boolML
if self.__boolML :
ordpoints=self.__midLine[:]
npoints=len(ordpoints)
xpoints=[point[0] for point in ordpoints]
ypoints=[point[1] for point in ordpoints]
polyOrd=PolygonRoi(xpoints, ypoints, npoints, PolygonRoi.POLYLINE)
return polyOrd
#if self.getMaxF()<15 : return None
#self.__FeretAxis()
#return self.__line
self.__boolML=True
self.__image.killRoi()
self.__image.setRoi(self.__contour)
boundRect=self.__contour.getBounds()
boundRoi=Roi(boundRect)
xori=boundRect.x
yori=boundRect.y
wori=boundRect.width
hori=boundRect.height
ip2 = ByteProcessor(self.__image.getWidth(), self.__image.getHeight())
ip2.setColor(255)
ip2.setRoi(self.__contour)
ip2.fill(self.__contour)
skmp=ImagePlus("ip2", ip2)
skmp.setRoi(xori-1,yori-1,wori+1,hori+1)
ip3=ip2.crop()
skmp3=ImagePlus("ip3", ip3)
skmp3.killRoi()
#-------------------------------------------------------------
if debug :
skmp3.show()
IJ.showMessage("imp3 l287")
#-------------------------------------------------------------
IJ.run(skmp3, "Skeletonize (2D/3D)", "")
#IJ.run(skmp3, "Skeletonize", "")
#-------------------------------------------------------------
if debug :
skmp3.show()
IJ.showMessage("imp3 l294")
#-------------------------------------------------------------
IJ.run(skmp3, "BinaryConnectivity ", "white")
ip3.setThreshold(3,4, ImageProcessor.BLACK_AND_WHITE_LUT)
IJ.run(skmp3, "Convert to Mask", "")
#-------------------------------------------------------------
if debug :
skmp3.show()
IJ.showMessage("imp3 l302")
#-------------------------------------------------------------
#IJ.run(skmp3, "Skeletonize", "")
#-------------------------------------------------------------
if debug :
skmp3.updateAndDraw()
skmp3.show()
IJ.showMessage("imp3 l308")
#-------------------------------------------------------------
rawPoints=[]
w=ip3.getWidth()
h=ip3.getHeight()
rawPoints=[(x+xori,y+yori,self.__sommeVals(x,y,ip3)) for x in range(w) for y in range(h) if ip3.getPixel(x,y)==255]
tempbouts=[val for val in rawPoints if val[2]==2]
if len(tempbouts)!=2 : return None
# test
#if len(tempbouts)!=2 :
#
# IJ.run(skmp3, "BinaryConnectivity ", "white")
# ip3.setThreshold(3,3, ImageProcessor.BLACK_AND_WHITE_LUT)
# IJ.run(skmp3, "Convert to Mask", "")
# #-------------------------------------------------------------
# if debug==debug :
# skmp3.updateAndDraw()
# skmp3.show()
# IJ.showMessage("if test l 328")
##-------------------------------------------------------------
# rawPoints=[(x+xori,y+yori,self.__sommeVals(x,y,ip3)) for x in range(w) for y in range(h) if ip3.getPixel(x,y)==255]
# tempbouts=[val for val in rawPoints if val[2]==2]
ip3.setRoi(boundRect)
if rawPoints==[]: return None
npoints=len(rawPoints)
xpoints=[point[0] for point in rawPoints]
ypoints=[point[1] for point in rawPoints]
valpoints=[point[2] for point in rawPoints]
bouts={}
if tempbouts==[]: return None
if tempbouts[0][1]>tempbouts[1][1]:
bouts["A"]=tempbouts[0]
bouts["B"]=tempbouts[1]
else:
bouts["A"]=tempbouts[1]
bouts["B"]=tempbouts[0]
rawPoints.remove(bouts["A"])
rawPoints.remove(bouts["B"])
rawPoints.append(bouts["B"])
tempList=[val for val in rawPoints]
p=bouts["A"]
Dist={}
ordPoints=[]
for j in range(len(rawPoints)):
Dist.clear()
for i in range(len(tempList)):
dx=p[0]-tempList[i][0]
dy=p[1]-tempList[i][1]
d=math.sqrt(dx*dx+dy*dy)
Dist[d]=tempList[i]
distList=Dist.keys()
mind=min(distList)
nextpoint=Dist[mind]
ordPoints.append(nextpoint)
tempList.remove(nextpoint)
p=nextpoint
ordPoints.insert(0, bouts["A"])
npoints=len(ordPoints)
if npoints < 4 : return None
xpoints=[point[0] for point in ordPoints]
ypoints=[point[1] for point in ordPoints]
polyOrd1=PolygonRoi(xpoints, ypoints, npoints, PolygonRoi.POLYLINE)
f=min(self.__midParams[0], len(xpoints)//2)
angleA1=polyOrd1.getAngle(xpoints[0],ypoints[0], xpoints[1],ypoints[1])
angleA2=polyOrd1.getAngle(xpoints[1],ypoints[1], xpoints[2],ypoints[3])
angleA = (angleA1+angleA2)/2.00
angleA=polyOrd1.getAngle(xpoints[0],ypoints[0], xpoints[f],ypoints[f])
angleA=angleA*(math.pi/180)
angleB1=polyOrd1.getAngle(xpoints[-2],ypoints[-2], xpoints[-1],ypoints[-1])
angleB2=polyOrd1.getAngle(xpoints[-3],ypoints[-3], xpoints[-2],ypoints[-2])
angleB = (angleB1+angleB2)/2.00
angleB=polyOrd1.getAngle(xpoints[-f],ypoints[-f], xpoints[-1],ypoints[-1])
angleB=angleB*(math.pi/180)
coef=self.__midParams[1]
xa = xpoints[0]-coef*f*math.cos(angleA)
ya = ypoints[0]+coef*f*math.sin(angleA)
xb = xpoints[-1]+coef*f*math.cos(angleB)
yb = ypoints[-1]-coef*f*math.sin(angleB)
lineA=Line(xpoints[0],ypoints[0], xa, ya)
lineB=Line(xpoints[-1],ypoints[-1], xb, yb)
lineA.setWidth(0)
lineB.setWidth(0)
lineA.setStrokeWidth(0)
lineB.setStrokeWidth(0)
ip2.setColor(0)
ip2.fill()
ip2.setColor(255)
ip2.setRoi(lineA)
lineA.drawPixels(ip2)
ip2.setRoi(lineB)
lineB.drawPixels(ip2)
ip2.setRoi(self.__contour)
ip2.setColor(0)
ip2.fillOutside(self.__contour)
ip2=ip2.crop()
imb=ImagePlus("new-ip2", ip2)
#-------------------------------------------------------------
if debug :
imb.show()
IJ.showMessage("imb l416")
#-------------------------------------------------------------
w2=ip2.getWidth()
h2=ip2.getHeight()
ip4 = ByteProcessor(w2+2, h2+2)
im4=ImagePlus("im4", ip4)
for i in range(w2):
for j in range(h2):
ip4.set(i+1,j+1,max([ip2.getPixel(i,j),ip3.getPixel(i,j)]))
#im4.show()
#-------------------------------------------------------------
if debug :
im4.show()
IJ.showMessage("im4 l430")
#-------------------------------------------------------------
im4.killRoi()
#IJ.run(im4, "Skeletonize (2D/3D)", "")
#IJ.run(skmp3, "Skeletonize", "")
#-------------------------------------------------------------
if debug :
imb.show()
IJ.showMessage("imb l300")
#-------------------------------------------------------------
#IJ.run(skmp3, "Skeletonize", "")
ip4=im4.getProcessor()
rawPoints2=[]
w4=ip4.getWidth()
h4=ip4.getHeight()
rawPoints2=[(x+xori-2,y+yori-2,self.__sommeVals(x,y,ip4)) for x in range(w4) for y in range(h4) if ip4.getPixel(x,y)==255]
self.__MidBouts=[val for val in rawPoints2 if val[2]==2]
# test
if len(self.__MidBouts)!=2 :
IJ.run(im4, "BinaryConnectivity ", "white")
ip4.setThreshold(3,3, ImageProcessor.BLACK_AND_WHITE_LUT)
IJ.run(im4, "Convert to Mask", "")
rawPoints2=[(x+xori-2,y+yori-2,self.__sommeVals(x,y,ip4)) for x in range(w4) for y in range(h4) if ip4.getPixel(x,y)==255]
self.__MidBouts=[val for val in rawPoints2 if val[2]==2]
ordpoints=[]
p0=self.__MidBouts[0]
rawPoints2.remove(p0)
c=0
while p0!=self.__MidBouts[1]:
if c<len(rawPoints2):
point=rawPoints2[c]
else: break
if abs(point[0]-p0[0])<2 and abs(point[1]-p0[1])<2:
p0=point
ordpoints.append(point)
rawPoints2.remove(point)
c=0
else: c=c+1
ordpoints.insert(0, self.__MidBouts[0])
self.__midLine=ordpoints[:]
self.__midCenters = self.__Centers(self.__midLine)
npoints=len(ordpoints)
xpoints=[point[0] for point in ordpoints]
ypoints=[point[1] for point in ordpoints]
polyOrd=PolygonRoi(xpoints, ypoints, npoints, PolygonRoi.POLYLINE)
#print self.__midLine
#print self.__MidBouts
#print xpoints
#print ypoints
return polyOrd
def main(imp,options):
from ij.plugin import ChannelSplitter
from ij.gui import Roi,PointRoi, PolygonRoi, Overlay, Line
from java.awt import Color
from ij import WindowManager
from ij.measure import ResultsTable
from ij.text import TextWindow
active_z=imp.getZ()
imps = ChannelSplitter.split(imp)
imp.setZ(active_z)
roi_int = imp.getRoi()
comp_imp=Zproj(imps[options["comp_ch"]],
"SUM",
active_z,
options["z_range"])
comp_imp=mode_subtract(comp_imp,roi_int)
loci_imp=Zproj(imps[options["loci_ch"]],
"SUM",
imp.getZ(),
options["z_range"])
loci_imp=mode_subtract(loci_imp,roi_int)
#Finding the boundaries of compartment and loci
comp_roi=thresh(sum_prj=comp_imp,thresh=options["comp_T"],roi=roi_int,method="boundary")
print "ok"
if (options["loci_method"]== "locus center"):
loci_roi=thresh(sum_prj=loci_imp,
thresh=options["loci_T"],
roi=roi_int,
method="point")
elif options["loci_method"]== "locus boundary":
loci_roi=thresh(sum_prj=loci_imp,
thresh=options["loci_T"],
roi=roi_int,
method="boundary")
if options["loci_method"]== "locus center":
dist,xc,yc,xl,yl=get_center_edge_dist(imp,comp_roi, loci_roi)
elif options["loci_method"]== "locus boundary":
dist,xc,yc,xl,yl=get_closest_points(imp,comp_roi,loci_roi)
rt_exist = WindowManager.getWindow("Loci distance to compartment")
if rt_exist==None or not isinstance(rt_exist, TextWindow):
table= ResultsTable()
else:
table = rt_exist.getTextPanel().getOrCreateResultsTable()
table.incrementCounter()
table.addValue("Label", imp.title)
table.addValue("Distance(micron)", dist)
if options['measure_feret']:
feret_roi,loci_feret,loci_area= feret(sum_prj=loci_imp,thresh=options["loci_T"],
roi=roi_int,pixel_size=imp.getCalibration().pixelWidth)
table.addValue("Loci feret", loci_feret)
table.addValue("Loci area", loci_area)
table.show("Loci distance to compartment")
## Adding loci overlay
ov=imp.getOverlay()
if ov==None:
ov=Overlay()
line = Line(xc,yc, xl,yl)
line.setStrokeWidth(0.2)
line.setStrokeColor(Color.PINK)
ov.add(line)
if options["loci_method"]== "locus center":
ov.add(PointRoi(loci_roi["x"],loci_roi["y"]))
elif options["loci_method"]== "locus boundary":
ov.add(loci_roi)
if options['measure_feret']:
ov.add(feret_roi)
ov.add(comp_roi)
imp.setOverlay(ov)
