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