Re: Doublet discrimination : Aria vs Vantage DiVa

I don't think there's a hard-and-fast rule about whether to use 
height vs. width on the Aria vs. Digital Vantage for doublets.	In my 
experience, both with a Digital Vantage and a FACSCanto, the 
parameters used to discriminated doublets depend more upon the cells 
than the instrument.

I start out with two dotplots, FSC-A vs. FSC-H, and SSC-A vs. SSC-W. 
Sometimes the H vs. A plot gives the best discrimination, sometimes 
the A vs. W plot is better, sometimes either one will do, and 
sometimes I use both, gating first through one, then further gating 
that singlet population through the second plot.

Try it out and see what works for you.	I think you'll find that cell 
morphology dictates your approach more than instrument choice.

Yoav Altman
Manager, High Throughput Cell Analysis
Burnham Institute for Medical Research
10901 North Torrey Pines Road
La Jolla, CA 92037


At 8:25 AM +0200 4/13/06, Roberts Joanna wrote:
>Hello there,
>Could anyone please offer me an explanation about doublet 
>discrimination in DiVa software
>on the Vantage versus on the Aria? ....inasmuch as i have believed 
>that on the Vantage
>setup in Diva, one uses FSC-A vs FSC-H to best acheive this while on 
>the Aria, width is
>used.
>Have I got the right end of the stick? Is it important?
>If I have got this right, could somone kindly explain to me why this is so.
>Thank-you so much,
>Joanna
>Flow Cytometry
>EPFL
>Lausanne
>Switzerland