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

[rocan <@t> mac.com]

Fat is notoriously difficult to cut on a cryostat.  At one time we 
"delipidated" the sample by leaving small pieces in acetone at 4°C; 
then we would freeze in OCT and cut.  More recently we embed in Cryogel 
from www.instrumedics.com and we do not need the acetone incubation 
anymore. Cryogel is a lot easier to cut than OCT. For mouse skin with 
fat this Cryogel make an enormous difference. A warmer temperature may 
be very helpful.  Think of it as if you were slicing frozen butter!
Also, we now also use instrumedics  tape transfer system called 
CryoJane for fat and it works particularly well but it is expensive and 
has a good learning curve before you get the best results. I only would 
buy this system if you are going to do fat sectioning on a regular 
basis.
I hope this helps.
Rocio

M. Rocio Sierra-Honigmann
Director
Engineered Wound Repair Laboratory
Cedars Sinai Research Institute
Tel.310-423-1882
Fax 310-423-2325
Honigmannr <@t> cshs.org

On Apr 20, 2004, at 7:52 AM, Nancy.Walker <@t> sanofi-synthelabo.com wrote:

> 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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