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looking for protocol using DAPI for cell cycle analysis - (Aug/31/2007 )


Have anyone had experience with DAPI staining for cell cycle analysis. Is it possible to co-staining other surfaces markers while using DAPI for cell cycle analysis. I have been trying to do cell cycle analysis in a rare population. But after sorting, i got only 10,000 cells , which are hard to use IP method for cell cycle analysis. I tried a commercial live dye which can co-staining with surface marker but it gave me very poor G0/G1 peak resolution. So, I really wish DAPI could give me some hopes.



well, DAPI staining have been performed and the cell cycle resolution looks very good. But my problem still unsolved blush.gif
DAPI staining is just like PI, except that DAPI does not need RNase.
DAPI is a better choice if we need to co-stain with surface marker, because PI overlap with many other channels, like PE ,APC and PE-CY7.
However, the results are confusing , despite the good cell cycle profile. I found no difference in cell cycle status in my Wild type and knock out cells. While I did see increased BrDu incoporation in Knock out cells.
Any suggestion?