Threshold problems

Posted by Leila Alieh on Oct 08, 2013; 6:56pm
URL: http://imagej.273.s1.nabble.com/Threshold-problems-tp5005085.html

Hello!

I developed a macro to count the number of cells positives for a given
antibody. The cells are selected depending on size, circularity and
intensity. I found an intensity value good for some images and I run the
macro. After that I found that the choosen threshold is too high for one
image, so that to have a reliable valuation for that image I should use
half of the threshold chosen for the other images, but this latter
threshold is really too low for the other images and it gives a wrong
result. I can't understand why this image is so different because the
intensity of the cells is not so different by eyes (it's a good quality
image, the ab signal is strong and there are no shadows). I was wondering
if I did some mistake during the "editing": I had to cut the images (I'm
interested only in a small area of them) and save them in a tiff format. Is
it possible that this editing chenged the intensity values?
Thank you for your help!!!!

Leila

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