def getMask(ip, method):
W = ip.getWidth()
H = ip.getHeight()
hist = ip.getHistogram(256)
stats = ip.getStatistics()
thresh = AutoThresholder().getThreshold(method, hist)
thresh = (thresh / float(255)) * stats.max
mask = ByteProcessor(W, H)
for i in range(W * H):
value = ip.getf(i)
bite = 255 if value >= thresh else 0
mask.set(i, bite)
fillHoles(mask)
mask.erode()
mask.dilate()
return mask
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
