Try Dexamethason 1 uM to make your thymocytes apoptotic.
After 3 hour of culture about 30% of your thymocytes are
Annexin V positive and PI negative. About 5% is positive for
both dyes.
When you do the incubation longer than 3 hr, you get more
apoptotic cells.
My suggestion is, make a time study an measure after every
hour the annexinV / PI . After this you can make your choice
what time you use in the future.
Succes
Cor Jacobs
Department of Nephrology
University Hospital, Nijmegen
Nijmegen, the netherlands
E-Mail: C.jacobs@nefro.azn.nl