Re: IMAGEJ Digest - 27 Jul 2010 to 28 Jul 2010 (#2010-35)

[Daniel James White]

Hi Ignacio,

On Jul 29, 2010, at 6:00 AM, IMAGEJ automatic digest system wrote:

> Date:    Wed, 28 Jul 2010 16:47:45 -0400
> From:    Ignacio Fernandez-Garcia <[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.

once you have the nuclear mask maybe you can dilate it to make it say 10 pix bigger then it will catch the centrosomes?


> 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?

possible, but might be a bit fiddly.... how to tell which is the right nucleus for each centrosome...?

better to keep it as simple as poss....

Dan


Dr. Daniel James White BSc. (Hons.) PhD
Senior Microscopist / Image Visualisation, Processing and Analysis
Light Microscopy and Image Processing Facilities
Max Planck Institute of Molecular Cell Biology and Genetics
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