Re: Help: Centrosome quantification

Posted by Bertrand Vernay on Jul 29, 2010; 7:20am
URL: http://imagej.273.s1.nabble.com/Help-Centrosome-quantification-tp3687435p3687436.html

Hi Ignacio

You may ant to have a look at this publication
PLoS Biol. 2008 Sep 16;6(9):e224.
A genome-wide RNAi screen to dissect centriole duplication and centrosome
maturation in Drosophila.

Dobbelaere J, Josué F, Suijkerbuijk S, Baum B, Tapon N, Raff J.
http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.00602
24

The authors used Cellprofiler (http://www.cellprofiler.org/index.shtml) to
score centrosomes defects. A similar protocol may work for you.

Regards

Bertrand
--
Dr Bertrand Vernay
Microscopy Senior Research Associate
Developmental Biology Unit
UCL Institute of Child Health
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-----Original Message-----
From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of
Ignacio Fernandez-Garcia
Sent: 28 July 2010 21:48
To: [hidden email]
Subject: Help: Centrosome quantification

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!