Hello,
I am trying to do a cell count for S2 cells from Drosophila. These are more or less adherent cells and I have to do a kill curve using puromycin. I did a trial run where I grew the cells in a 12-well plate, each with a coverslip, so the cells adhere and grow on the coverslip. This is so that, after each time point I can collect the coverslip, fix the cells and stain with a fluorescent marker like DAPI, and estimate live cells.
The fundamental assumption here is that, after the cell dies, it loses its adherent property and will eventually fall off the coverslip, and hence only the live cell population will remain on a coverslip, which I can fix and count.
The issue is, even towards very late time points, like 72 hours or 96 hours, I can see lice cells in cultures where I used extremely high puromycin, like 12ug/ml. The count of blue spots does go down on the 1st and 2nd day, but suddenly there is an increase towards later days.
Any suggestions?
Additionally, has anyone worked with S2 cells and know whether they lose the adherent property when they die? or they just keep sticking to the surface?
- Badges
- Science topic
- Similar topics
- Philosophy
- Philosophy Of Science
- Reasoning