Re: DAB quantification

Posted by Gabriel Landini on
URL: http://imagej.273.s1.nabble.com/DAB-quantification-tp5006159p5006164.html

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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