Hi Nan, If all you are looking for is cell death then I think you might try plotting PI against forward scatter. What you will probably see is a lot of PI positive stuff which you can infer has nuclei and maybe some low scatter material that is PI negative which would probably be debris. You could also try combining PI with FDA (fluorescein diacetate) staining where the live cells should cleave the FDA to form fluorescein so they will be green. I normally add 10ul FDA (1ug/ml) to 1ml cells and incubate for 10mins before adding PI. Derek >Dear flowers: > >One of the researcher in our lab wants to see the percentage of live or >dead cells after exposing cells to anoxic condition and after drug >treatment. I was just using PI to indicate dying cells for him. But I >think by doing this we are eliminating dead cell that had broken into >pieces. On the other hand, if I'm counting the debries (which are low >FSC and SSC), I am afraid that the live cells that are very small will >be treated as debries ,too. I am attaching the histogram of PI staining >and FSCvs.SSC here. R1 is what I considered live gate. I'm not sure what >I should call R2. And R4 is the PI positive gate. I was thinking >counting live cells is a better approach, but I don't know if there is >any marker conmercially available for this purpose. Would you please >give me some insight? >By the way, the cells that they use are C2C12 myoblasts, which the size >difference among cells is obvious. -- *************************************************************** Derek Davies Voice: (44) 020 7269 3394 FACS Laboratory, FAX: (44) 020 7269 3100 London Research Institute, e_mail: derek.davies@cancer.org.uk Cancer Research UK mobile: 07790 604112 44 Lincolns Inn Fields, London, UK. Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html In tenebris lux ***************************************************************Received on Mon Feb 9 15:38:00 2004
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