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confocal assay for FITC-labled peptide in Hela cell - (Apr/12/2006 )

Hi, I am fremman in confocal assay for Hela treated with FITC-labled peptide.
when i did, no cleaned FITC peptide often appeared, so there are always some FITC peptides aroud membrane. How can I solve this problem?
And i can not seperate the dead and live cell. the live cells will be easier to be pepentrated by FITC peptide or dead cells.
thank you!



have you incubate the cells at 37C?
how long have you incubate them and at which concentration?
did you used serum during incubation with the peptide?
if you need some nice result you can permeabilize transiently the cells during incubation...