TUNEL Questions.

Manuel Saenz Terrazas jmsaenz at UACHIH.UACHNET.MX
Wed Apr 2 17:15:04 EST 1997


Hi.
I am currently trying to perform a TUNEL test on thymus and spleen sections 
but i have troubles with the adherence to the slide glass. I prepared the 
mounting glass with 3-amino-propyltriethoxysilane which is said to improve 
the glass adherence specially for immunocytochemistry. The protocol 
somebody gave me is very simple: Wash the glass slide in 'silane solution 
(5% on pure acetone) for five minutes and let stand on soft paper until 
dry. However, after dewaxing the spleen sections i have only the stromal 
ring still attached to the glass and nothing more.I would like to know if 
somebody knows a better protocol to avoid this problem.

On the same issue, i am aware that FACS is today's standard for the measure 
of apoptosis, but a flow cytometer is several thousand dollars away from 
me. The question worrying me is the following: Does TUNEL replaces 
EtBr/AcrOr staining of cell suspensions for apoptosis index determination?

Thanks in advance:

Manuel




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