[Histonet] Autofluorescence problem on atheroma - TUNEL staining

[delphine eberle]

Hi,

 

Autofluorescence in atheroma is a well known problem for immunofluorescence staining.

I would love to hear input about how to reduce it to a minimum (if possible!).

 

So far, we are trying to detect apoptotic cells in atheroma by TUNEL staining (Roche, TMRred) and we found the interpretation very difficult as some area of the tissue show red dots, tat could be considered as apoptotic bodies. We are working with frozen sections, fixing with Formalin, quenching with Glycine etc... 

 

-I was wondering if acetone fixation (or other fixation methods) could decrease atheroma autofluorescence by extracting lipids and others materials? 

-Also, I am not sure that acetone fixation is recommended for TUNEL staining? Did anybody try?

-Are there around any approved ways to decrease autofluorescence in atheroma? (I can not find anything!)

 

Thanks!

 

Delphine

Delphine Eberlé PhD 
UCSF Department of Vascular Surgery
VA Medical Center - NCIRE
Building 2 - room 410
4150 Clement Street 
San Francisco, CA 94121, USA
Tel: 415 221 4810 ext.2984
Cell: 857 453 0821
delphine.eberle <@t> ucsfmedctr.org