[Cytometry] multiple negatives for comp in Diva?

Rupak Neupane rneupane at cellerant.com
Thu Feb 6 12:56:48 EST 2014


You don't mention what your acquisition software is, but FACSDiva allows you to have tube specific negative control. All you have to do is add a gate for negative population (I use the auto-interval gate). If a tube has both a positive and a negative gate, then the software uses that negative, if it doesn't have a negative gate, it uses the universal negative from unstained tube. So, at least on Diva you won't have that problem. Other acquisition software could work differently though. Hope that helps.
Rupak.


Rupak Neupane
Manger, Flow Cytometry Core Facility
Cellerant Therapeutics, Inc.
1531 Industrial Rd
San Carlos, CA 94070
650-232-5437
rneupane at cellerant.com


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Carol Norris
Sent: Thursday, February 06, 2014 8:40 AM
To: cytometry at lists.purdue.edu
Subject: [Cytometry] multiple negatives for comp in Diva?

Hi Flow Gurus,

We have lots of folks combining GFP-expressing cells with antibody labeling, and I'd like to get more of them using comp beads for the antibody controls. There doesn't seem to be a way to use GFP-negative cells for the GFP comp control and unstained beads for the antibody comp controls in Diva.  Are unlabeled cells OK to use as a universal negative?  The other option would be to transfer the data to FlowJo to get a comp matrix, and then transfer the comp values back into Diva.

Thanks,
Carol


Carol E. Norris, Ph.D
Facility Scientist
Flow Cytometry/Confocal Microscopy Facility Biotechnology/Bioservices Center University of Connecticut Unit 3149
91 N. Eagleville Rd
Storrs, CT 06269-3149

Phone (860) 486-3080
Fax (860) 486-5005




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