Unfortunately the HTS software (now part of Diva) on the LSR II does not allow you to delay the acquisition. That's a (would be) a very important feature for many reasons and it is available on the Macintosh for the Calibur. The reason of the disturbance is too high cell concentration and/or presence of many RBCs. It is a hydrodynamic disturbance that settles after a couple of seconds of run. In the absence of acquisition delay you should dilute your samples and try to eliminate RBSs as much as possible. We hope BD is polishing up the HTS software on Diva. Especially because I am about to order three of them. Akos ________________________________________________ Akos Szilvasi Flow Cytometry and Confocal Microscopy Core Facilities Novartis Institutes for Biomedical Research, Inc. (NIBRI) 100 Technology Square, Cambridge, MA 02139 akos.szilvasi@novartis.com 617 871 7177 office 617 871 7051 fax "Poon, CK" <cpoon@medsfgh.ucsf.edu> 05/19/2005 05:22 PM To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> cc: (bcc: Akos Szilvasi/PH/Novartis) Subject: RE: The Mystery of The High Throughput Sampler, The Sampler Port, and the Clumped Cells Hello, We use our HTS with the LSRII here at UCSF. We have done about 15 runs (PBMCs) on our HTS and have not experienced any clogging. However, I did notice that with our concentration of cells ( ~8million PBMC/ml), the dots start to jump around on the plots if we run faster than 1.5ul/s (a sign of clogging?). I also noticed that keeping the TUBE/PLATE rocker switch in the TUBE postion when running the HTS tends to screw the run up. I don't know if this will help and I don't know anything about the boost pressure. Good Luck, Ck Poon Research Associate Core Immunology Lab UCSF/SFGH -----Original Message----- From: Villasmil, Rafael [mailto:VillasmilR@MedImmune.com] Sent: Wed 05/18/2005 02:32 PM To: cyto-inbox Subject: The Mystery of The High Throughput Sampler, The Sampler Port, and the Clumped Cells Good Afternoon Everyone I want to find out why the sample port clogs when I use the HTS but it works fine when doing Manual Tube Loading (MTL) I have an assay that runs perfectly when I use tubes to load the samples. However, the Calibur sample port clogs constantly when I use the HTS. Looking at the samples under the microscope, we found that there are some clumped cells. We think that the boost pressure of the HTS is higher than the boost pressure of the Calibur when doing MTL Any comments are welcome Thanks Rafael Villasmil Associate Scientist I Medimmune Inc Phone Number (301) 398-4478 One MedImmune Way Gaithersburg, MD 20878 "Accept the challenges, so that you may feel the exhilaration of victory." George S. Patton<?xml:namespace prefix = o ns = "urn:schemas-microsoft-com:office:office" />Received on Mon May 23 17:46:11 2005
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