Hello,
I normally usually crystal violet to measure cell viability, but the cell lines have all been adherent. I need to measure viability with suspension lines post siRNA transfection. I was thinking of using poly ornithine to fix them to 96-well plates and then use a live and dead stain to take time lapse images using incucyte. Now I'm wondering if the fixed cells might detach as they die? Not sure if the fixation process is permanent or not. If they do detach as they die like adherent cells then I could just fix them, wash away the dead ones after treatment and use my normal crystal violet method. Anyone have any insight, experience, suggestions? Thanks!
I would recommend to use MTT assay to stain such cells. But with some modification - I was adding MTT solution 10x more concentrated that usually used (to avoid medium removal), then after incubation (2-3h) centrifuge your plate 5min 1200 RPM in room temparature. Then they should be attached to the bottom of the plate, so you can safely remove the liquid and dissolve the formazan crystals. I was using this method for HL-60 and KL562 cells. Best luck!
I agree. MTT is so well but in some cases (polycyclic compounds for example) they could give false positive results. You can use double check in some experiments to compare MTT and other one. More, you can stain cells using Trypan Blue solution, then wash them and count stained/unstained cell using any cell counter or hemocytometer.
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