FRET

Randy T. Fischer fischer1 at mail.nih.gov
Tue Jan 16 13:43:17 EST 2007


Leonie,

Why not tune your primary laser to 458 and your second laser to 568.  You
can successfully trigger off of scatter using 458 and still use it for the
CFP.

Randy T. Fischer, NIH/NIAMS
B Cell Biology Group
9000 Rockville Pike
Bldg 10, Room 6D50
Bethesda, MD 20892
(301) 594-3537 (voice)
(301) 402-2209 (fax)


> From: Leonie Gaudry <L.gaudry at unsw.edu.au>
> Date: Tue, 16 Jan 2007 10:36:38 +1100
> To: Cytometry Mailing List <cytometry at flowcyt.cyto.purdue.edu>
> Subject: FRET
> 
> 
> Hi,
> I have been asked to do FRET using mCherry to GFP and also CFP to YFP.
> I was going to tune my second Laser to first 458nm for CFP then 568
> for the mCherry.
> Are there any alignment beads to confirm detection at these
> excitation wavelengths as wells as emission  at 475nm for CFP anf
> 610nm for the mCherry?
> As we are looking for energy transfer I presume I would have to turn
> off the 488nm Laser. Does this mean  triggering on fluorescence for
> the mCherry and CFP?
> I would appreciate any help from the community.
> Thank you
> Leonie Gaudry
> 




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