Nancy Perlmutter ngp at shapirolab.com
Mon Feb 23 11:38:19 EST 2004

Mark Coles wrote:
>I am interested in analyzing lymphocytes that express DsRed.  I could 
>analyze the samples on a FACS calibur but I have noticed that both the 
>excitation and emission spectrum of DsRed is not very compatible with the 
>calibur.  Has anybody had success with the analysis of low levels of DsRed 
>expression on the calibur or other instrumentation.   Has anyone tried a 
>machine that has a green laser to excite DsRed?  We have a luminex machine 
>which has a green laser, is it possible to use that machine to analyze cells?

Yes the Luminex-100 instrument can be used to analyze cells.  I have used 
the Luminex instrument to analyze peripheral blood stained for CD4, CD8, 
and CD45 with various combinations of PE, CY5, APC, APC-CY7.

However to use the instrument for cell analysis you will need to increase 
the gain on the APDs (used to detect the red emitting fluorochromes) and 
possibly adjust the PMT (used to detect PE) voltage. You will also need to 
adjust (lower) the threshold level for the doublet discriminator (side 
scatter). To make these changes to the instrument settings you will need 
access to the Luminex Maintenance menus. The analysis of the data should be 
done using third party FCS compatible software. I use FCS Express. The 
original Luminex software wrote "somewhat" FCS 2.0 compliant files.

If you or anyone else is interested in using the Luminex instrument for 
cell analysis please contact me directly and I can supply more details.


Nancy Perlmutter ngp at shapirolab.com
Laboratory Manager
Howard M. Shapiro, MD, PC
Cytometry Laboratory
119 Braintree Street, Suite 102
Allston, MA 02134
phone 617-783-8392  fax 617-783-4750

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