Posted by
Joel Sheffield on
URL: http://imagej.273.s1.nabble.com/Quantifying-Stained-Retina-tp5000880p5000996.html
Hi All,
First, the stain that Natalie is referring to is known in the histo world
as "Perls Prussian Blue". A brief description follows:
a stain for ferric iron as in hemosiderins, using potassium ferrocyanide in
acetic acid or dilute hydrochloric acid followed by a red counterstain such
as safranin O or neutral red; various hemosiderins and most mineral irons
give a blue-green reaction, while nuclei stain red.
Note that the procedure often includes a counterstain. Natalie, do you
know if your procedure includes some other dye, such as those mentioned
above? If so, that would account for Gabriel's concerns about red in the
image. Being color blind, I see very little of that color on my monitor,
but there can be lots of variation in monitors. I will make the
assumption, for the sake of argument, that there actually is a
counterstain, in which case I would recommend that you try the stain
without it. It might be possible to resolve the contribution of the Perls
stain under those conditions.
One more point, since you are interested in layers, it will be much easier
to quantify the stain per layer if you take your images so that the layers
are horizontal.
Joel
On Wed, Nov 28, 2012 at 5:07 PM, Gabriel Landini <
[hidden email]>wrote:
> On Wednesday 28 Nov 2012 20:55:11 Natalia Chacon wrote:
> > I've enclosed another image that seems to contain more iron in the
> sample.
> > Also, is it still possible to use the Color Deconvolution to get the
> > data my mentor desires?
>
> Hi, I do not think there is a lot of blue in your images, so I doubt that
> it
> can be done reliably.
> Look at the colour cube of your image (color inspector 3d) and you will
> realise what I mean.
>
> Cheers
>
> Gabriel
>
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--
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
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