RE: Sorting Issues

From: Nebe-Von-Caron, G (g.nebe-von-caron@unipath.com)
Date: Thu Aug 15 2002 - 07:01:04 EST


You can calculate your dilution factor of sample volume by taking into
consideration your sample flow (for example 18 ul per minute) and your
droplet generation  (for example 30 kHz)per minute. You therefore
transfere a sample volume of 18ul / 1800000 per droplet = 10fl,
independent of pressure settings. Depending on your target volume into
which you sort you can then calculate molarities.

The sample volume flow is dependent on the differential pressure between
sheath and sample, the viscosity of your sample, the resistence of
tubing... easiest just measured by weighing the tube before and after a
run w/o boosting sample flow and dripping flow back into the sample
tube.

Regards

Gerhard



-----Original Message-----
From: Lee, Siow Fong [mailto:sflee@cha.ab.ca]
Sent: 12 August 2002 19:23
To: cyto-inbox
Subject: Sorting Issues



Hi All,

	I am posting the following questions for a colleague
(cmattar@ualberta.ca) who uses a Beckman Coulter Ultra for sorting:

	When sorting an event from a post-nuclear cell supernatant,
during
the actual sort when the event is diverted into the sorting vessel, does
some of the cell lysate that the event was suspended in originally also
enter the sorting tube?  Will soluble proteins floating around in the
post-nuclear supernatant get sorted along with the event of interest?  I
realize the sample is taken up along with sheath fluid.  Does this also
dilute the non-specific soluble proteins which may contaminate a sorted
sample?  Thanks.

Siow Fong Lee
Department of Laboratory Medicine & Pathology
University of Alberta Hospital
Capital Health Authority
Edmonton, Alberta
Phone: (780)407-8067
Fax:  (780)407-8599
E-mail:  sflee@cha.ab.ca

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