Protocol Online logo
Top : New Forum Archives (2009-): : Flow Cytometry

Dot plot cell cycle analysis-strange dots - (Jan/28/2011 )

hi everyone,
i am doing cell cycle analysis via hoechst and pyronin Y. I can distinguish apoptotic cells, G1, G0 and S/G2/M. but I also have dots in my dot plot at higher hoechst intensities than the S/G2/M-population. what is it?
thanks in advance!

-ija0-

Did you gate-out doublets and aggregates?

-Denis Baev-

yes, SSC-W vs. SSC-A

-ija0-

It is not enough.
To precisely gate-out all "strange guys" you have to do the same on FLx-W vs FLx-A (FLx is an FL channel there you detect the DNA dye signal).

-Denis Baev-

ok, than I'll try this. thanks!

-ija0-