[Histonet] IHC with H & E staining

[Amos Brooks]

Hi,
    I usually try to avoid eosin as a counterstain for a DAB labeled slide
because the red/pink of the eosin can obscure the rusty brown of DAB. If
you really want to use it though I would suggest a *really* light eosin,
perhaps even just a few milliliters in the 95% ETOH as you are dehydrating
it. The RBCs and eosinophils will pick it up quickly and the stain should
not overwhelm the DAB. You could also darken the DAB with Copper sulfate.

Cheers,
Amos

On Wed, Jan 6, 2016 at 1:00 PM, <histonet-request at lists.utsouthwestern.edu>
wrote:

> Message: 6
> Date: Wed, 6 Jan 2016 11:26:35 +0000
> From: Marcus Green <marcus.green at oncology.ox.ac.uk>
> To: "histonet at lists.utsouthwestern.edu"
>         <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] IHC with H & E staining
> Message-ID:
>         <669331A24821FE4491665E8DFD47CF22C254A4 at MBX06.ad.oak.ox.ac.uk>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Dear Users,
>
>
>
> Happy New year - I hope this finds everybody well!
>
>
>
> I was asked a very simple question yesterday - why don't you do H&E
> counterstaining on DAB stained samples. The question came about as we're
> looking at CD31 staining for blood-vessels and some look ruptured (we're
> keen to see red blood cells).
>
>
>
> Instead of doing a consecutive cuts, a CD31 stain (Haem counterstain) on
> one slide and H&E on another, the question was asked why not do the Eosin
> stain as part of the counterstain....
>
>
>
> I've never seen it done in the literature or in the clinic, and I've never
> asked why it's not done. Any assistance or advice would be greatly welcome
> - and my sincerest apologies if this is a very basic question?
>
>
>
> Thanks in advance for your time,
>
> kind regards,
>
>
>
> Marcus,
>