Re: Quantifying Stained Retina

Posted by NatyC on
URL: http://imagej.273.s1.nabble.com/Quantifying-Stained-Retina-tp5000880p5001008.html

Dear Joel,

Unfortunetly we can't go back and restain these images.
The staining was done outside of our lab by a three party it seems.
So that means I'll have to work with what I'm being provided.

Joel you mentioned this might assist me with research.

"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."

However, how can I go about doing that?
Do I have to go to the orignal sample and start from there, or is
there a plugin I can use to assist me with this process?

Thanks,
Natalia



On 11/28/12, JOEL B. SHEFFIELD <[hidden email]> wrote:

> 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
>>
>> --
>> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>>
>
>
>
> --
>
>
> Joel B. Sheffield, Ph.D
> Department of Biology
> Temple University
> Philadelphia, PA 19122
> Voice: 215 204 8839
> e-mail: [hidden email]
> URL:  http://astro.temple.edu/~jbs
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>

--
ImageJ mailing list: http://imagej.nih.gov/ij/list.html