A means by which dead cells can be discriminated from live cells, when
both are labeled with CFSE, is to use 7-AAD. Only dead cells are permeable
to 7-AAD and its emission peak is far enough removed from the broad range
of CFSE emission to be easily compensated.
Kevin G. Waddick, Ph.D.
Parker Hughes Institute
tessa kerre wrote:
> Dear flowers,
>
> In addition to Dr. Waddick's questions concerning CFSE, we too have some
> problems with it.
>
> I have been using CFSE to track human cord blood HSC in vivo and check
> for cell divisions.
>
> - I cannot use frozen material, because then I get clump forming, which
> results in loss of 75% of my cells. Is this normal?
> - It is extremely difficult to compensate the overflow from CFSE towards
> the FL-3 and the FL-2 channel, making it difficult top combine CFSE with
> PE or Cy labeled antibodies. Gating out of dead cells using propidium
> iodide also becomes very difficult. Do others also have the same
> problems, or do you have the magical settings?
> - How long can you keep CFSE in good shape frozen in DMSO (-20°C)?
>
> Kind regards,
>
> Tessa
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