[Histonet] processing adipose tissue
I have adipose tissue to process (fix and paraffin embedding) for in situ
hybridization (ISH) and IHC experiments. I've done this in the past with a
program and got pretty good tissue morphology and done some experiments,
but since I was told by Gayle Callis that my processing schedules for
rodent brain and embryo are probably excessively long. In fact in a recent
experiment where I included an embryo slide prepared by Novagen; Signals
were much better with the commercially supplied slide.
So does anyone have any advice on processing fat? I'm using small samples
of epididymal fat (0.5g - 1 g per sample ) that have been fixed overnight
in PFA 4% and washed in PBS for 6h.
This is the program we used :
alcohol 70% 1h
paraffine 3X1h30 (Sakura tissue tech III)
(Our embedding machine does have possibilities to do vacuum and heat).
Before doing IHC or ISH we just deparaffin with xylene (2x5') and
rehydrate. We do Protein K before in situ and microwave or trypsin or EDTA
before IHC . Are there some specific pretreatments for adipose
tissue for delipidizing that might improve probe or antibody accessibility.
I appreciate your advice and thanks for your valuable time,
Molecular Biology Scientist
B.P. 37 Labége Innopole
31676 LABEGE CEDEX FRANCE
tel : (33)561004179 fax :(33)561004001
Histonet mailing list
<< Previous Message | Next Message >>