Re: [ Alexa selection question ]

Hi,

Alexa 700 can't be read at FL3. (Alexa 700 needs red excitation; FL3 is a 
blue PMT)

635 will excite Ax700 but you cannot read it in FL4 as latter has a BP 
filter around 660

You can use either Ax 647 or  APC in a single tube and read it in FL4.

PE-Cy5 (FL3) and APC (FL4) can be used simulataneously in FACSCalibur. 
Experiment needs to be carefully designed to minimise compensation.
Pls refere to Purdue email archives for previous discussion on this.

regards
Paresh


Dr. Paresh Jain
Scientific Advisor / Technical Services

BD Biosciences
Signature Tower-B, South City 1, Gurgaon, Haryana 122001 India 
tel: +91 124 2383566  cell: +91 9811275447  fax: +91 124 2383224/5/6
E-mail: paresh_jain@bd.com   Website: www.bd.com




RAJA ALAMELU <alameluraja@yahoo.com> 
01/05/2007 04:37 PM

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Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
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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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