[Histonet] Double fluorescent stains for apoptosis

From:"Chan Wai Kam"


We’re doing a study on the effect of compression and degeneration of rabbit intervertebral discs, and we're using the In Situ Cell Death Detection Kit, TMR red (Roche) for the detection of apoptosis by fluorescent microscopy.  As our lab is new to this technique, we would be grateful for any advice on the correct apoptosis stain/kits to use.  

Our problem in using this kit is that we get a large number of positive stained cells (all red).  As we would like to find out the percentage of apoptotic cells, we need a protocol which can give us a double stain which according to one of the papers is bright green (tunnel positive) and light red (unaffected cells).  

Any advice and protocols would be greatly appreciated.

Thanks in advance
Julee Chan
Orthopaedic Surgery
Histology Lab
National University of Singapore 

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