if Annexin-v and PI staining is suitable to attached cells - (Oct/27/2007 )

now i am try to use Annexin-V and PI double staining to see if the drug which now i am using could induce the attached cells go to apoptosis,
but i met some problems that it is very difficult for me me to decide when i should stop trypsiniation in order not to hurt the cells

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alternatively to flow cytometry, you can utilize fluorescence plate reader; cells must be grown on fluorescence plate reader usable cell culture plates (cytowell f.i.); or another alternative is inspection by confocal microscopy and counting of events; in each alternative, you do not detach cells;

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:)staining


thank you so much for your help

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Bearer said:

alternatively to flow cytometry, you can utilize fluorescence plate reader; cells must be grown on fluorescence plate reader usable cell culture plates (cytowell f.i.); or another alternative is inspection by confocal microscopy and counting of events; in each alternative, you do not detach cells;

How is the background when you use fluorescence plate reader? Annexin/PI not bound to cells need to be discarted?

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