[Histonet] frozen lung sections

[Tony Henwood]

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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