counting cells in multiclour fluorescence images

Posted by Matthias Kirsch on Nov 16, 2008; 8:58pm
URL: http://imagej.273.s1.nabble.com/counting-cells-in-multiclour-fluorescence-images-tp3694463.html

Dear all,

I am trying to find a way to count cells expressing one or the other or both proteins in cultured cells (see e.g. the attached example). There are green cells, red cells and yellow cells and I would like to have a (semi)-automatic routine with which this can be done objectively. Basically the cells can be identified by their nuclei (in the blue channel), but I do not know how to count only the nuclei which have a red, green or yellow cytoplasm associated with them. As there probably is no way to reliably find the outline of a cell with any accuracy (however, maybe maybe one has an ingenious approach), it may be sufficient to look in the immediate vicinity of the nuclei (e.g. in a ring some pixels wide around the nuclei -user selectable preferrentially), if this area is red, green or yellow.
I would appreciate if someone could help to solve this problem.

Thanx
Matthias