4 replies to this topic

[acute]

Dear All,
does any one have suggestion how to detect dapi with flow cytometry? should it be one plot (FL4) or two plot (SSC/ FL4)?
can flow cytometry detect the apoptotic bodies using dapi staining?
many thanks

[vraev]

It would depend on the cytometer that you are using. I use the LSR2 and there is a DAPI parameter installed in the BD software. I just use DAPI/SSC-A to select cell populations.

cheers,

V

[acute]

vraev, on Nov 4 2009, 12:04 PM, said:

It would depend on the cytometer that you are using. I use the LSR2 and there is a DAPI parameter installed in the BD software. I just use DAPI/SSC-A to select cell populations.

cheers,

V

thanks but how do you analyse your cell population? when apoptotic bodies formed in a population, what should I see?
of course there is a rise in SSC but what about FL4?

[vraev]

well...I don't do intensive analysis with DAPI. I just look at the positively stained population in comparison with an FMO for DAPI and that way I can look at the dead cell percentage in my cell preperation. thats all I use it for... to see how much of my cells are dead.

[blotted]

vraev, on Nov 6 2009, 06:40 AM, said:

well...I don't do intensive analysis with DAPI. I just look at the positively stained population in comparison with an FMO for DAPI and that way I can look at the dead cell percentage in my cell preperation. thats all I use it for... to see how much of my cells are dead.

you can see apoptotic cells quantifying subG1 peak. For this purpose I recomend you to use PI.