[Histonet] frozen lung sections
Tony Henwood
AnthonyH <@t> chw.edu.au
Wed Apr 7 20:16:31 CDT 2010
Betsy,
I would dissolve the OCT in Hanks solution (or similar cell culture
medium - not saline). Place in dilute OCT (50/50 with distilled water),
place on stirrer until the lung settles in the OCT 30-60min. Blot with
absorbent paper, refreeze in fresh OCT. It should cut quite well.
I have used this previously on some tissue that was received the same
way. It seems that the OCT had "freeze-dried" prior to receipt in my
lab.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
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-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Molinari, Betsy
Sent: Thursday, 8 April 2010 12:46 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] frozen lung sections
Hi all,
I have received some lung samples on OCT. These lungs have not been in
PBS or any fixative. They went directly from dissection into OCT and
frozen.
I cannot get a section. The temp of the cryostat is -20. I have done
some reading and it seems the best results come from immersion in PBS,
then 4% paraformaldehyde then into sucrose. Can anyone clarify the
procedure or offer advice? I am unfamiliar with this technique since
most of my frozen sectioning is heart tissue and it does not present the
same problem as the lung.
Thanks,
Betsy
Betsy Molinari HT(ASCP)
Texas Heart Institute
Cardiovascular Pathology
6770 Bertner Ave
MC1-283
Houston,TX 77030-2607
832-355-6524
832-355-6812
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