[Cytometry] multiple negatives for comp in Diva?

Will Schott Will.Schott at jax.org
Tue Feb 11 09:47:28 EST 2014

You can also delete the P2 histogram gate and make a new P2 or P2 and P3
on a dot plot instead as long as the P2 and P3 are subsets of the P1
scatter gate. I have had to do this a few times when users gave me samples
that had more than one stain in them (like PI and GFP).

William Schott
Flow Cytometrist
The Jackson Laboratory
600 Main St.
Bar Harbor, ME 04609
Phone: (207) 288-6192

On 2/6/14 12:40 PM, "Cris Bare" <flowmail at verizon.net> wrote:

>It's easier than you think.
>In DiVa on the comp controls worksheet in the fluorochrome specific
>histogram where you adjust gate P2 to select the positive population, you
>can draw a P3 gate on the negative population and DiVa will use P3
>preferentially for calculation.
>That's a word sentence.
>1. Include negatives in each comp control tube.
>2. Add a P3 gate on the negatives in the same histogram as P2 positives.
>3. Calculate. 
>This is why DiVa has the checkbox when you create your comp tubes to
>include or not include a universal negative.
>On Feb 6, 2014, at 8:40 AM, Carol Norris <carol.norris at uconn.edu> wrote:
>> Hi Flow Gurus,
>> We have lots of folks combining GFP-expressing cells with antibody
>>labeling, and I¹d like to get more of them using comp beads for the
>>antibody controls. There doesn¹t 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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