[Histonet] fat cryostat sections: cutting pb, IHC?

[Alan Bright]

Nancy,

The sample you are trying to section should not be a problem, it looks to me that your knife temperature and anti-roll plate are not cold enough, you omitted this temperature in your email perhaps you could inform me. Also you may need to have the specimen slightly warmer. I hope this assists you.

Best Regards

Alan Bright

Bright Instrument Co.Ltd.
St Margaret's Way
Huntingdon
Cambridgeshire
PE29 6EU
England

Tel No:+44 (0)1480 454528
Fax No:+44 (0)1480 456031
Email: abright <@t> brightinstruments.com
Web Site: www.brightinstruments.com



-----Original Message-----
From: Nancy.Walker <@t> sanofi-synthelabo.com [mailto:Nancy.Walker <@t> sanofi-synthelabo.com] 
Sent: 20 April 2004 15:53
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] fat cryostat sections: cutting pb, IHC?


Any pointers for cutting adipose tissue with a cryostat?

 I have  mouse epididyme adipose tissue that is unfixed and embedded  in tissue tek. At - 30°C I can cut  without it melting but most of the specimen stays on the knife edge leaving a nicely cut section of tissue tek, a big hole and a little fat tissue around the edges. And this at 45 µ!!!  Any thinner and I get just the hole.

I have a crostat which I can adjust the block and chamber temp separately. Haven't fiddled with the knife angle yet...

WHAT TO DO???

any special tratments to get IHC reagents to penetrate this hydrophobic stuff?

thank you for you advice,

Nancy Walker
Molecular Pharmacology

Sanofi-Synthelbo Research
B.P. 37 Labége Innopole
31676 LABEGE CEDEX FRANCE

nancy.walker <@t> sanofi-synthelabo.com
tel : (33)561004179  fax :(33)561004001




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