Date: Thu, 04 Jan 2001 09:27:17 +0000
From: "Mike Loughlin" <Mike.Loughlin@sheffch-tr.trent.nhs.uk>
To: "Greer, Bonnie" <Bonnie.Greer@stjude.org>
Subject: Re: RED BLOTCHES IN GOMORI`S TRICHROME?
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This is a common artefact found in this technique.  The following might be =
helpful:
Recommended technique
1) Air dry cryostat sections for only 2 minutes.
2) Stain in modified Harris's haematoxylin for 5 minutes (Filter stain =
before use and check pH to exactly 2.3 before use)
3) Very quickly rinse in 3 changes of distilled water.
4) Stain in modified Gomori's stain for 10 minutes.
5) Drain slide and wipe off excess stain from the back of the slide and =
round the section on the front.
6) Agitate the slide in a coplin jar containing 0.2% acetic acid for 1 =
second only
7) Immediately dehydrate very rapidly through graded alcohols to xylene =
and mount in synthetic mountant.

NOTES
1) The pH must be kept low throughout the procedure
2) Dyes must be pure, Sigma Fast Green FCF and Chromotrope 2R recommended.
3) The two dye solutions may  be prepared and stored in the fridge.  Once =
the two dye solutions have been combined, the solution can be stored in =
the fridge and used again but discarded as soon as staining becomes pale.

I assume that you have details of the staining solutions but let me know =
if not.  I hope following the above will give you even sharp staining.  If =
not, if you could send me details of your exact technique I will try to =
help further.  GOOD LUCK!!
Mike Loughlin
Histopathology Department
Sheffield Children's Hospital
UK


>>> "Greer, Bonnie" <Bonnie.Greer@stjude.org> 01/03/01 09:04pm >>>
Help why  does this happen..  does a guru know the answer...=20