Help: Centrosome quantification

Posted by Ignacio Fernandez-Garcia on Jul 28, 2010; 8:47pm
URL: http://imagej.273.s1.nabble.com/Help-Centrosome-quantification-tp3687435.html

Hi all,
Lately I've been working analyzing the centrosome abnormalities in some experiments. One thing we're interested in is the number of centrosomes per cell (usually two in normal cells). To detect the centrosomes we use an immunoflurescence technique where we label the centrosomes in green and counterstain the nuclei with DAPI in blue. So, we obtain images like this one:

http://a.imageshack.us/img715/4948/tubulincaexample.jpg

As you can see, we have some green signals (the punctual ones) distributed within the cell cytoplasm (seen as green background) but not necessarily within the nucleus. This way, a nuclear mask with the DAPI is not enough to segment the centrosomes and quantify them as a "number-of-centrosomes-per-cell" value.
One approach that comes to my mind is: Segment both, centrosomes and nucleus, in their correspondent channel, and then associate the segmented centrosome signals with the nearest segmented nucleus by the minimum square distance between mass centers.
So, my question is, anybody can help me with that? does anybody have done it before?
Also I'll really appreciate suggestions, comments, questions, etc.
Thanks!
Cheers!