FACS Aria _very high compensation

From: Wijewardana, Viskam <viskam@pitt.edu>
Date: Thu Jul 10 2008 - 15:39:24 EDT
Hi,
I have a compensation problem. I use Pacific Blue, FITC, PE, PerCP and APC for my Antibody panels and I use FACS Aria 1 for analysis. I do compensation using BD CompBeads according to BD protocol and do auto compensation in FACS Diva. My problem is I get very high compensation due to bleeding as shown below.

PerCP-PE 70.35%
APC-PerCP 102.83%

Other seems OK. I use following detectors and lasers as set by our flow manager.

405 Pacific Blue 450/50
488 FITC 530/30 with 505LP
488 PE 575/25 with 550LP
488 PerCP 710/50 with 685LP
633 APC 660/20

I tried, for PerCP the 660/20 with 635LP in the blue laser, then it looked worse.
Any suggestions?

Thanks
Viskam

Viskam Wijewardana
Center for Vaccine Research
9051 Biomedical Sciences Tower 3
University of Pittsburgh
3501 Fifth Avenue
Pittsburgh, Pennsylvania 15261
Ph 412 383 9057
Fax 412 624 4577
Received on Fri Jul 11 14:18:00 2008

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