Hello,

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.

-Tobias-


________________________________________________________________
Verschicken Sie romantische, coole und witzige Bilder per SMS!
Jetzt neu bei WEB.DE FreeMail: http://freemail.web.de/?mc=021193


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet