RE: [Histonet] Frozen embedding of lungs

From:"Louro, Pedro"

Hi Kim,

I have done this procedure before with good results.
I used a 50:50 ratio of OCT and saline mixed well together into a
syringe, then inflate the lungs with this mixture.
I tried to use all OCT but it was impossible to inflate the lungs
because the OCT was so thick.  
After inflation with the 50:50 ratio, I froze the lungs in OCT either of
two ways: 

1) Plastic embedding mold with OCT into Methyl Butane that is cooled
with liquid nitrogen OR
2) Metal embedding mold with OCT into a slushy mix consisting of dry ice
and Methyl Butane.

Both give good results 
Hope this helps

Pedro Louro
Schering-Plough Research Institute

-----Original Message-----
From: Kim O'Sullivan [] 
Sent: Wednesday, March 29, 2006 7:47 PM
Subject: [Histonet] Frozen embedding of lungs

Hi all,

Does any one out there have a successful method for inflating mouse
lungs to freeze. We currently freeze them unfixed in OCT, but of course
the morphology is shocking as the lungs have collapsed. Is there a
method that preserves the morphololgy without collapsing or imploding
the lungs.

Any advice will be appreciated

Kim O'Sullivan
Centre for Inflammatory Diseases
Department of Medicine
Monash University

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