[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 4°C 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 10µm thick.
Thank you for your help.
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