96 well FACS

Laird Bloom LBloom at Phylos.com
Tue Oct 9 10:19:18 EST 2001


	Hi,
	The only way around this event limit that I've found is to use the
threshold settings.  The Cytek unit has a connection directly to the
cytometer's threshold circuitry, and if you give CellQuest a high value for
threshold in FSC and FL1, for example, you can get rid of both small
particles and cells of low FL1 fluorescence.  I've used this successfully a
few times.  Unfortunately, you don't save any of the data points that fall
below the threshold, so you can't reconstruct what you've gated out.  But at
least you can collect up to 63,000 events that fall above the thresholds
you've set.
	Maybe Cytek can comment on this question--a higher event limit and a
longer sampling time (currently under 90 sec.) would be really nice.
Laird Bloom
> Phylos, Inc.
> 128 Spring Street
> Lexington, MA  02421
> lbloom at phylos.com
>
> ----------
> From:		Woda, Marcia
> Sent:		Tuesday, October 9, 2001 10:51 AM
> To:	'Laird Bloom'
> Subject:	RE: 96 well FACS
>
> can you get around this 63,000 event limit???
>
> marcia woda-umass worcester facs
>
> -----Original Message-----
> From: Laird Bloom [mailto:LBloom at Phylos.com]
> Sent: Monday, October 08, 2001 9:41 AM
> To: Cytometry Mailing List
> Subject: RE: 96 well FACS
>
>
>
> Hi,
>	I have not used the BD loader, but my lab has had a 96-well
> autoloader from Cytek Development for about 6 months, and we have been
> very
> satisfied with it.   It can process a plate in about 40 minutes and can
> handle volumes down to 80-100 ul.  The cost is around $15,000, which is
> much
> cheaper than what BD is offering.  The Cytek unit has an awkward interface
> with CellQuest and can't adjust its acquisition based on events falling
> within a particular gate, and so it might not be useful for people who
> have
> to analyze rare cell populations.  But for routine screening or other
> applications where acquiring up to 63,000 events will give you enough of
> the
> events you want to allow subsequent analysis in CellQuest, I would highly
> recommend it.  It certainly has extended our capacity to do large-scale
> experiments with a small number of people.
>	Contact information for Cytek:
> Cytek Development:	510-657-0102.  Website http://www.cytekdev.com/
> I arranged a demonstration through Ray Lannigan at Tritech Field
> Engineering, 800-886-7004
>	I submitted a more detailed review to this e-mail list in June.
> Please contact me directly if you cannot retrieve it from the archives and
> would like a copy.
>
> Laird Bloom
>
> Phylos, Inc.
> 128 Spring Street
> Lexington, MA  02421
> lbloom at phylos.com
>
>
>
>
>
>
> > ----------
> > From:		Ellen.Freed at astrazeneca.com
> > Sent:		Thursday, October 4, 2001 3:17 PM
> > To: Cytometry Mailing List
> > Subject:	96 well FACS
> >
> >
> > Hi All
> >
> > Has anyone yet used the 96 well format from BD FACScalibur.  How is it
> > working?  Is 96 well format available from anyone else?  Have not kept
> > track
> > of other conversations along this line.  Thanks
> >
> > Ellen Freed
> >
> > Ellen Freed
> > Scientist
> > AstraZeneca R&D Boston
> >
>



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