[Histonet] too blue

[Amos Brooks]

Hi Stacy,
     Trichrome stains are REALLY sensitive to fixation variations. I have a
great image of two heart samples stained simultaneously. Both were formalin
fixed, but one was fixed overnight and the other (if you can call it fixed)
was dropped into a full container and probably saw a half hour fixation
total, and since it was on top of a pile of cassettes probably never really
was immersed. The properly fixed specimen has red muscle and blue collagen
as is expected in heart tissue. The other cut miserably, has folds &
wrinkles, and the muscle is mostly light blue with darker blue collagen. Red
blood cells are red, and some blushes of muscle fibers have taken on some
faint red, but the difference is striking. Fixation is probably more
critical to proper staining than the actual time in staining solutions. That
is where I would start looking.

Good luck,
Amos


Message: 3
Date: Fri, 25 Jun 2010 13:52:27 -0400
From: "Stacy McLaughlin" <Stacy_McLaughlin <@t> cooley-dickinson.org>
Subject: [Histonet] too blue
To: <histonet <@t> lists.utsouthwestern.edu>
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Any ideas why patient tissue stained with Trichrome would be too blue when
the controls are working perfectly?
We've repeated the stain and it keeps happening.
Thanks

Stacy McLaughlin HT(ASCP)
Lead Histology Tech./Laboratory Safety