Re: Intensity quantification area

Posted by Gabriel Landini on Oct 31, 2013; 11:12am
URL: http://imagej.273.s1.nabble.com/Intensity-quantification-area-tp5005401p5005403.html

On Thursday 31 Oct 2013 10:40:29 Rbalada wrote:

> So, I'm trying to quantify the intensity of an Immuno Histo Quemistry from
> rat hippocampus and I'd like to measure it using always the same rectangle,
> with the same exact area, so then I can compare the different intensities
> regarded in each picture from each condition. That's my first doubt, how do
> I get sure of using always the same rectangle?
>
> Second question: how can I correlate each intensity point within a rectangle
> with the exact point of the picture? (the coordenates). For what I've been
> told, ImageJ homogenizes the different intensity points within a rectangle,
> so you can not know something like 'at the coordenate x= 0,5 and y= 1,3 the
> intensity is...whatever. So, is that like this or there is an option to see
> each intensity value within a rectangle? If so, how could I do this? Any
> idea of how could I know this intensity values using some other program if
> ImageJ does always homogenize the intensity within a rectangle? Maybe the
> homogenized intensity within a rectangle may work for comparing between
> conditions, but not between different points within the same rectangle of
> the same condition.

Unfortunately it is not possible to quantify immunohistochemistry reliably.
See the "Note" in red here:
http://www.dentistry.bham.ac.uk/landinig/software/cdeconv/cdeconv.html

You might be able to capture some intensity of stain, but that would not tell
reliably much about how much product there is present which relates to the
amount of antigen present.

To be able to quantify the amount of products, you need a stoichiometric
reaction (IHC is not so) AND a chromogen that follows Beers-Lambert law.

Cheers

Gabriel

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