Re: Resorufin

From: Kelly Hardwicke (kellyh@ccmi.salk.edu)
Date: Wed Jul 19 2000 - 12:56:15 EST


Hello Artem -

Just an idea - you may have already considered this but your message does not
mention it.

Have you brought down the FSC threshold? (Ours is usually set at the 72nd
channel where at most, if not all FSC gains something as small as E. coli would
lost).  If your flow cytometer is set to threshold at a higher FSC channel than
the E. coli will occupy the machine won't let on that it's seeing them, no
matter how bright they actually are!

I would also try to align the instrument to optimally detect the smallest beads
you can find.

Another thing to try would be thresholding on the fluorescence channel as
opposed to the  forward scatter channel.

Good luck,
Kelly Hardwicke
Salk Institute Center for Cytometry and Molecular Imaging, Assistant

On Tue, 18 Jul 2000, Artem Khlebnikov wrote:
>Hello all,
>
>I have question in regard of 2-dodecyl resorufin form ImaGene Red C12RG
>lacZ kit (Molecular probes).
>
>I can see stained E. coli under the microscope and detect fluorescence on
>fluoremeter... no problem!!
>and it is specific to cells
>but have no signal on FACS or used 513, 568 and 590 excitation lines
>and bunch of different filters.
>
>Does any one has experience with it?
>Thanks a lot in advance,
>
>Artem



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