FW: sections lifting...

From:jim

Michelle

We routinely work with brain tissue from human, sheep, rat, mouse, etc. The
trickiest would have to be our human double hemisphere slices that we do
both routine H/E and even immunohistochemistry. Once the section is picked
up on the glass slide (150mm x 120mm slides are used)it is left to drain in
a vertical position. The slides can be left for a few minutes and are
blotted between at least 2 layers of moistened Whatmans filter paper using a
roller. The slides are then placed in racks and loaded onto a heated blower
for no more than 10 minutes. This has never failed for doing routine stains
like the H/E, Weil etc. The glass slides are cleaned by the lab staff and
are not coated at all. If doing harsher special stains such as
Immunohistochemistry we then need to polysine coat the slides.

Cheers

Jim Manavis
Senior Hospital Scientist
Department of Neuropathology
Institute of Medical & Veterinary Science
Adelaide, SA, 5000
Australia
Phone: 61-08-8222-3668
FAX: 61-08-8222 3204
e.mail: jim.manavis@imvs.sa.gov.au
Disclaimer: Not this little black duck!


-----Original Message-----
From: Michelle Mcanulty-Smith - NeuronZ Ltd.
[mailto:Michelle.Mcanulty-Smith@neuronz.com]
Sent: Monday, 9 September 2002 12:39 PM
To: 'histonet@pathology.swmed.edu'
Subject: sections lifting...


Hi Folks,

I have been asked to post a question to  histo land and hope that someone
may be able to help...

We have whole adult rat brain (60 day rats approx 300gm weight of the brain)
the brains are perfused with Saline and fixed in 4% Formalin. The brain is
left to fix in for 48 hours minimum and then processed (cycle 16 hours using
chloroform)

The brains look and feel great after processing and cut beautifully... all
is perfect until.... the sections float off when staining! It has been
suggested to me that this is a processing problem.

We have tried various and almost all slide adhesives from pre-coated to
gelatine coated. We have even tried different kind of gelatine as I am told
pig is the best.

I have suggested drying for longer in our 58 degree oven even overnight, but
to no avail which is why 'they' now think that it is a processing problem.

The morphology of the bits that do stay on the slide is great,  we are
staining with Thionin and Acid Fuchsin and other people are not experiencing
the problem.

All thoughts and suggestions most welcome.
Michelle

Michelle McAnulty-Smith
Histology Co-ordinator

NeuronZ Ltd
Auckland
New Zealand
Tel: 09 367 7167 ext. 7060
Fax: 09 367 7186

E-mail:  michelle.mcanulty-smith@neuronz.com


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