I see a similar discrepancy when I sort bacteria (SYTO9 stained) with a FACSCalibur (bacterial suspension is checked by microscopy to ensure there is no clumping). However, I get excellent accuracy sorting green Calibrite beads. Any additional thoughts on why a Calibur may struggle to report correct numbers of sorted bacteria, even though purity of sorted bacteria is usually ~99%, would also be appreciated. Daniel Hoefel PhD Student University of South Australia CRC for Water Quality and Treatment, Australian Water Quality Centre Private Mail Bag 3, Salisbury SA 5108, Australia Ph: +61 8 8259 0325 Fax: +61 8 8259 0228 Email: daniel.hoefel@sawater.com.au -----Original Message----- From: McDonald, Ken [mailto:ken_mcdonald@Merck.com] Sent: Thursday, 28 November 2002 9:12 AM To: cyto-inbox Subject: Vantage cell counts Hi All, I'm seeing a discrepancy in what the Vantage sorter tells me it's sorted and the actual numbers sorted. For example, I sorted for green calibite beads (30,000) and when I re-ran them to check for purity there were roughly 15,000 only. Does anyone have the same problem? The purity is good (99%) and everything else checks out by the book?..Thanks in advance..Ken *******South Australian Water Corporation disclaimer******* "This e-mail and any attachments to it may be confidential and/or subject to legal professional privilege. If you are not the intended recipient you may not disclose or use the information contained in the message in any way. If received in error please delete all copies and contact the sender by return e-mail. No warranty is made that any attachments are free from viruses. It is the recipient's responsibility to establish its own protection against viruses and other damage."
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