[ Alexa selection question ]

Randy T. Fischer fischer1 at mail.nih.gov
Fri Jan 5 14:41:37 EST 2007


Alamelu,

For the FACSCalibur, you would be best off using a tandem for FL3,
preferably a PE-Cy5.5, PerCP-Cy5.5, or a PE-Alexa Fluor 700.  Try to not use
PE-Cy5 as there is an immense spectral overlap with APC in the FL4 channel.
And unless you have made some very interesting modifications of your
Calibur, there is only one channel off of the red laser and that is the APC
equivalent FL4 (Alexa 647 can be run here but then no APC).

Good luck,

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: RAJA ALAMELU <alameluraja at yahoo.com>
Date: Fri, 5 Jan 2007 03:07:46 -0800 (PST)
To: cyto-inbox
Subject: [ Alexa selection question ]

Dear Flow list members,
We have a FACS Calibur with 2 lasers - 488 and ~635nm and emission optics to
measure FL1, FL2, FL3 and FL4.
Can we use Alexa Flour 700 as FL3? When viewing the Spectrum at BD site,
Alexa Fluor is falling totally within the range of FL3 emission filter
without any overlap with FL2 or FL4.
But will the 635 laser be able to excite Alexa700?
Will Alexa 647 be useful, instead of Alexa700 and without interference with
FL4 APC?
Will PE-Cy5 be useful, instead of Alexa700 and without interference with FL4
APC?
Those who have experience with FACS Caliber, kindly give your suggestions
please.
With thanks,
Alamelu Raja
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