From: Josep M. Gasol (pepgasol@cucafera.icm.csic.es)
Date: Thu Aug 06 1998 - 03:50:33 EST
Dear friends, Following some recent comments I would like to ask if anybody knows for sure what's the "true" rate of particles that the electronics of a FacsCalibur can see without losing events due to them passing in the dead time zone. My experience counting natural planktonic bacteria suggests that a flow rate above 500 particles per second underestimates true concentration (measured with an alternative method). This can certainly be due to two bacteria coming in the same drop through the laser, but could also be due to the electronics not being able to handle that many events per second. By the way, anyone has an idea of a way of checking for two particles going in the same drop ? Increasing bead concentration and relating doublets to singlets could be a way. Anyone ever tried ? A further related question: If I am gating what I see with a "software gate", (I mean not an electronic gate), I imagine that I should be concerned with the "total flow rate" (particles in my gate and those outside the gate) and not with the "acquired data rate". Am I right ? Thanks for your help ! Josep "Pep" Gasol ************************************************************ Josep M. Gasol Institut de Cičncies del Mar, CSIC Pg. Joan de Borbó, s/n E-08039 Barcelona, CATALUNYA Spain Phone: 34 93 221 6416 Fax: 34 93 221 7340 email address: pepgasol@icm.csic.es Department's web page: <http://www.icm.csic.es/bio/index_bio.html> ************************************************************
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