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 _____________________________________________________________________ Disclaimer This email and any files transmitted with it, is confidential and intended solely for the use of the individual to whom it was addressed. Unipath monitors emails to ensure its' systems operate effectively and to minimise the risk of viruses. Whilst it has taken reasonable steps to scan this email, it does not accept liability for any virus that may be contained in it. _____________________________________________________________________
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