Re: 2 queries

From:Shirley Powell


Xylene is still the best clearing agent for processing tissues.

I personally use positive charged slides here, but there are occasions when I
need to use an additive in the water bath.  I have used the commercial additive
Sta-On from Surgipath.  But many years ago when everyone was using Gelatin in
their waterbaths, which gave me a terrible bluish background staining, I found a
procedure in an old, very old histology text by Ann Preece (which is no longer
in print) for an agar solution to use in the waterbath.  My ancient records show
that it was a  .25gm agar agar (the kind they used in the bacti lab) to 100 mL
of distilled water and dissolve with the aid of heat.  Add a preservative, the
procedure calls for a grain of thymol (Thimerisol) to prevent contamination.  If
my memory serves me right, the solution gels, but it can be emulsified using a
blender.  Add 20 mL of agar per 1000 mL of water depending on the size of the
waterbath.  I still use agar at times and keep it in the frig and shake it well
before using it because it gets little thick when cold.

For your brain sections, make sure all the water has drained from between the
paraffin section and the slide by drying them in a vertical position before
heating and staining.

Agustín Venzano wrote:

> Dear Netters: I'm needing two opinions from you:
> 1. What are your thoughts and experience on the use of chloroform as a
> clearing agent when processing nervous tissues?
> 2. We are currently picking up brain sections on slides that haven't been
> previously coated with any adhesives. Some time ago we used Mayer's albumin,
> but we gave up this one due to its disgusting physical characteristics. What
> kind of adhesive substances are you presently using for routine H&E
> histopathology?
> Thank you in advance
> Agustin Jose Venzano Halliburton
> DVM- INTA Castelar, Argentina

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