Re: Rare event sorting....

Hello Chris
You've left out lots of primary information about cell type, machines etc so
here are 2 quick responses to you:

First what machine are you sorting on?	Older analog systems may have less
efficiencies to sort such rare pop'ns and indeed you could very well be
grabbing unwanted stuff.  Newer sorters have really good efficiencies.
Also, make sure as many parameters (ie antibodies, dead cell identifiers
etc) as possible are used to truly identify the rare pop.
Second, if the cells you are trying to attain are involved in a
developmental assay, put them in culture and see if biologically they do
what they are supposed to do.  The usual procedure will be to culture these
cells for a while and then review them with standard facs.  The reviewing
usually tells you if you got the right ones or not.
Ok hope this helps somewhat
> I've got a question.	I've got some folks that are needing to sort out small
> populations.
> .1% or so and smaller.
> 
> How do people prove to themselves that what they've got coming out is what
> they want when
> you're looking at a sorted quantity of 500 cells or so?  At least
> that's what they expect to get in some cases.
> 
> I'd like to be sure we're actually doing what we think we're doing.
> 
> 
> Thanks,
> Chris
> 
> 
> 
> 

-- 
Gisele Knowles
Cytometry and Scanning Microscopy Core Facility
Sunnybrook and Women's Research Institute
B wing 210
Sunnybrook Health Sciences Centre
2075 Bayview Avenue, Toronto, ON
M4N 3M5
Ph: 416- 480-6100 x 7282, 7284
Fax: 416-480-4375