[Histonet] Help with mouse adipose tissue
Kinsley, David
david.kinsley <@t> spcorp.com
Wed Mar 1 12:32:04 CST 2006
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
908-740-4669
*********************************************************************
This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete.
More information about the Histonet
mailing list