[Histonet] Autofluorescence problem in olfactory epithelia cryosections

From:=?iso-8859-1?Q? "Tobias=20M=E4tzig" ?=


I've got a serious autoflourescence problem when trying to perform 
ISH on mouse head cryosections.

After ISH process I can see a bright red autoflourescence in large 
parts of my tissue even without any antibodies on it.
First I thought it might be due to blocking solution containing Horse serum, 
but I test blocked my slides and there was almost no autoflourescence at all. 
So were the untreated slide !

Right now, I don't have a single clue which step of my ISH might cause
the red autofluorescence but it would be great if anyone of you had a good=20
advice for me. Please, let me know !

The mouse has been killed, decapitated, parts of the nose were
removed and fixed in 4%PFA for 3h. After that the tissue was 
incubated at 4C in 30% succrose until the tissue sank to the 
bottom of the tube and was than shock frozen and embedded in 
mounting medium. Slices are 10m thick.

Thank you for your help.


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