Re: Help: Centrosome quantification

Posted by Crowell Elizabeth on Jul 29, 2010; 10:24am
URL: http://imagej.273.s1.nabble.com/Help-Centrosome-quantification-tp3687435p3687438.html

Hello Ignacio,

This might be a pretty dumb question, but are you sure you need to
associate each centrosome with a nucleus?  If you are just interested in
the number of centrosomes per cell, why not segment both, count the
total number of nuclei per image, count the total number of centrosomes
per image, and determine the ratio?
Of course it would not give you an exact value, but if you simply want
to show statistical differences between conditions, it might be all you
need to do.
If you are worried about counting cells that are on the image borders
and whose centrosomes might not be included in the image, there are ways
to set filters in the "analyze particles" function to avoid counting
nuclei that are only partially visible.  I think your error rate would
be fairly low and you would get a very good estimate of the number of
centrosomes per cell.  Maybe worth testing...

Best regards,
Elizabeth


Ignacio Fernandez-Garcia a écrit :

> 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!
>  

--

Elizabeth CROWELL

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