I am trying to analyse the membrane intensity in the bleb and the cell body (please refer to the PDF attached). I need to plot the normalized intensity with the length of the bleb (more like dynamics of the bleb). Could some one please help me with how to analyse it. I have been trying to use QuimP plugin, but it is only giving me out mean intensity values of the whole cell.
kRAS mCherry.tif (6.3 MB) Doc1.pdf (47.0 KB)
I believe - regarding QuimP - the best person to answer your specific questions is @Piotr_Baniukiewicz.
If your blebs are also pretty well ‘blebbed’… ie - you have two more-or-less circular objects… you can always separate them using watershed. So something like this (using the image you shared):
//run("Threshold...");
setAutoThreshold("Otsu dark");
run("Create Mask");
run("Erode");
run("Dilate");
run("Fill Holes");
run("Watershed");
run("Analyze Particles...", "size=1000-Infinity display exclude add");
Output looks like:
And then you can measure what you need once you have those two ROIs saved… maybe that could work?
