Re: supporting tissue while paraffin embedding

From:Geoff McAuliffe

Hi Janet:

    You will need something that shrinks at the same rate as the tissue during
processing/dehydration. I think your choices are a good start. Maybe mix in some
albumin? Don't worry about GFAP staining, it is very resistant to processing (I
use DAKO rabbit anti-cow GFAP at 1:5000 with a Vector ABC Elite kit and
nickel+cobalt intensification). While isolectin B4 does stain microglia, it is
not specific, it also stains small, unmyelinated fibers in the spinal cord and a
few other things that escape me at the moment. I don't know if it works on rat
FFPE sections, I do mice. You migh try OX-42 and some of the ED antibodies
(ED-1) for rat microglia.
See J. Comp. Neurol. 314:125-135, 1991 and J. Histochem. Cytochem. 41:481-487,

"Miller, Janet" wrote:

> Hi, I was wondering if someone had any advice for me.  I am working with
> hydrocephalic rat brains, which have enlarged ventricles- making them
> essentially a balloon.  I was wondering if anyone had some advice as to what
> I can inject in to the ventricles after cardiac perfusion and a short post
> fix with 4% paraformeldehyde solution, that would help support the tissue
> during Paraffin processing.  I am currently working with GFAP and Isolectin
> B-4 microglial stains, so I would prefer if what ever method of support did
> not interfere with staining.  I was thinking of one of 3 different ways I
> could achieve this.
> 1) Agar
> 2) gelatin
> 3) HistoGel
> If anyone has any thoughts on these or any other suggestions, and
> concentrations I could try of the different materials, I would appreciate
> it!
> Thank you very much.
> Janet

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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