[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).
-will


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:

>Carol,
>
>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.
>
>Cheers!
>-cbb
>
>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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>
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