[Histonet] Help with mouse adipose tissue

From:"Kinsley, David"

Hi everyone,

I'm working on a project that involves collecting adipose tissue from
multiple sites of a mouse for the purpose of IHC.
I fixed my samples in 4% paraformaldehyde for 22 hours, rinsed in tap water
to remove excess formalin, then put them into 70% ETOH for 10 days and then
processed on my routine mouse program.
(30 min each in 80%, 95%, 95%, 100%, 100%, 100% ETOH then 30 min. each in 3
changes of Xylene, then 1 hour each in 3 changes of paraffin.)

I test sectioned 2 of the blocks to make sure they sectioned well.  The
first block was a smaller piece of fat from between the abdominal muscles
and skin.  It sectioned well and even stayed intact on a 50 degree water
bath.  The second piece (larger in size) from the epidydimal area was more
difficult to cut, but I was able to float it on a room temp water bath and
obtain sections good enough for H&E staining.  When I sectioned both blocks
again 2 days later, the smaller piece still cut well, but the larger piece
did not cut.  I tried ice with no water, and even freezing spray but both
did not help.

Does anyone have suggestions?  If I need to reprocess what would be the best
procedure to follow since these are for IHC?

thanks in advance

David Kinsley
Schering-Plough Research Institute
Kenilworth, NJ 07033

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