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quantification of DAB staining

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quantification of DAB staining

rona baron
27 posts
Hi all,

I stained tissues with antibodies specific for neurons (MAP-2) and with
hematoxylin which stains all the nucleus population, than imaged it with
light microscope.
Is it possible to distinguish between the brown staining for neurons and
the purple staining for the nucleus?
I would like to quantify the number of brown particles that I have out of
the total purple particles....


Thanks,
Rona

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Re: quantification of DAB staining

carlo bianco
28 posts
hi
you can try the color deconvolution way ( landini's plugin)
or the color segmentation,
moreover, is available online a software developed for ki-67 nuclear staining
quantification http://breast-cancer-research.com/content/12/4/R56
maybe at 40 x magnification it could work.
Let me know
Good luck

carlo  bologna Italy




________________________________
 Da: rona baron <[hidden email]>
A: [hidden email]
Inviato: Martedì 12 Marzo 2013 10:20
Oggetto: quantification of DAB staining
 
Hi all,

I stained tissues with antibodies specific for neurons (MAP-2) and with
hematoxylin which stains all the nucleus population, than imaged it with
light microscope.
Is it possible to distinguish between the brown staining for neurons and
the purple staining for the nucleus?
I would like to quantify the number of brown particles that I have out of
the total purple particles....


Thanks,
Rona

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Re: quantification of DAB staining

Shall, Sydney
8 posts
In reply to this post by rona baron
Dear Rona,
There is a plug-in for ImageJ crafted by Dr Pawel Znojek, which
essentially aims to do this.
He constructed it for DAPI and a fluorescent  antibody to a DNA repair
protein.
You should be able to adapt this easily for your purposes.
The plugin is available from the following site:
http://www.pzfociez.com/ .
I would recommend that you should add a plugin constructed by Gabriel
Landini, at the University of Birmingham, England, which helps considerably.
This you can find on the ImageJ site.

If you proceed down this path, I should be very grateful to you if you
could let me know how things develop for you.

With best wishes,

Sydney



On 12/03/2013 09:20, rona baron wrote:

> Hi all,
>
> I stained tissues with antibodies specific for neurons (MAP-2) and with
> hematoxylin which stains all the nucleus population, than imaged it with
> light microscope.
> Is it possible to distinguish between the brown staining for neurons and
> the purple staining for the nucleus?
> I would like to quantify the number of brown particles that I have out of
> the total purple particles....
>
>
> Thanks,
> Rona
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>


--
Professor Sydney Shall,
Department of Haematological Medicine,
King's College London,
Medical School,
123 Coldharbour Lane,
LONDON SE5 9NU,
Tel & Fax: +44 (0)207 848 5902,
E-Mail: sydney.shall,
[correspondents outside the College should add; @kcl.ac.uk]
www.kcl.ac.uk


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