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DAPI and Flow Cytometry


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#1 acute

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Posted 03 November 2009 - 09:16 AM

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

#2 vraev

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Posted 03 November 2009 - 11:34 PM

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. :lol:

cheers,

V

#3 acute

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Posted 04 November 2009 - 12:34 AM

View Postvraev, 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?

#4 vraev

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Posted 05 November 2009 - 09:40 PM

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.

#5 blotted

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Posted 22 December 2009 - 05:17 AM

View Postvraev, 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.





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