Jennifer,
Looking at the images, is it possible that the slides were air-dried for too
long prior to staining?
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Laboratory Manager Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer Fricton
Sent: Wednesday, 1 June 2011 2:28 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help with Masson's Trichrome - not working on cryo sections
Hello,
I am completely stumped by a problem we¹ve been having with our Masson¹s
Trichrome. It isn¹t working properly on cryo sections. We developed our
protocol several years ago and it has been a standard in our lab without any
trouble, until about four months ago when it just stopped working on frozen
sections. It still works fine on paraffin sections.
You can see images at this link:
http://www.med.umn.edu/lhi/prod/groups/med/@pub/@med/@dom/@lhi/documents/con
tent/med_content_340494.pdf
What seems to be happening is that the beibrich scarlet + acid fuchsin reagent
is either not staining the muscle tissue, or is washing out of the tissue. The
aniline blue is still staining the connective tissue very well (it stopped
working, too, but we solved by reducing time in water rinse following stain).
My Masson¹s protocol is very standard essentially straight out of the Armed
Forces Institute of Pathology Manual (p. 94). I fix my cryosections in 10% NBF
x 30 mins. before starting the protocol.
I¹ve made sure my formalin is fresh and properly stored, with no white
precipitate. I¹ve tried the Bouin¹s mordant step at both 56C and room temp
overnight. I¹ve reduced my distilled water rinses following the biebrich
scarlet and aniline blue steps to just a quick dip, thinking maybe I was
de-staining too much before setting the stains with acetic acid. I¹ve re-made
all solutions, checked all components and pH levels. I haven¹t done anything
different with my frozen tissues, and I¹ve tried several different freshly cut
tissues.
Has anyone seen this before and have any tips on what is going wrong? Your
help is greatly appreciated!
--
Jennifer L. Fricton, B.S.
Scientist, Lab Manager
Lillehei Heart Institute Histology Microscopy Core Facility University of
Minnesota School of Medicine, Division of Cardiology
312 Church Street SE
4-290 Nils Hasselmo Hall
Minneapolis, MN 55455
Lab: 612-626-3090
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
*
This email and any files transmitted with it are confidential and intended
solely for the use of the individual or entity to whom they are addressed. If
you are not the intended recipient, please delete it and notify the sender.
Views expressed in this message and any attachments are those of the individual
sender, and are not necessarily the views of The Children's Hospital at Westmead
This note also confirms that this email message has been virus scanned and
although no computer viruses were detected, The Childrens Hospital at Westmead
accepts no liability for any consequential damage resulting from email
containing computer viruses.
*
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
