Posted by
Panayiota Ploutarchou on
URL: http://imagej.273.s1.nabble.com/DAB-quantification-tp5006159p5006165.html
Hi Gabriel,
Thanks for your answer. I agree that I can't quantify, but doesn't darker (and more) DAB staining correlate to more antigen present? I was just wondering whether there is some sort of semi-quantitative way to go about doing this...
Many thanks for your help,
Yiota
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From: ImageJ Interest Group [
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Sent: Friday, January 17, 2014 4:19 PM
To:
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Subject: Re: DAB quantification
On Friday 17 Jan 2014 15:42:22 Panayiota Ploutarchou wrote:
> So, I've done Immunohistochemistry on tissue sections, and finished with DAB
> staining (which basically gives brown staining at the locations where a
> reaction with the antibody took place). Now, I would like to quantify that,
> but the pixel intensity function of Image J seems to give weird results
Unfortunately IHC is not stoichiometric and so you won't get a reliable
measure of how much product you have. Most stains are not "quantitative" (with
some exceptions like Feulgen and phalloidin for example).
> and also how would you suggest I go about quantifying
> the DAB staining as I would need to compare staining in control and treated
> sections.
You can't, immunostains are not a quantitative technique. What you see as dark
has no accurate relation to the amount of antigen present, which is what the
purpose of the quantification is.
Cheers
Gabriel
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