We've noticed a strange, but substantial quirk on our DiVa systems that we're wondering if others could corroborate. The symptom presented itself as a large discrepancy in the number of Threshold events (# of events recognized by the instrument) versus the number of events Processed (# of events that actually get processed and saved in your file). More specifically, the more events you collected (Threshold) the fewer events were actually saved (Processed). Typically, the reason this occurs is because of coincidence aborts. Since there is no dead time on DiVa instruments, the abort rate is usually only about 1%, assuming you have your window extension set properly. But, what we were seeing is that the processed event count 'plus' the abort count, which should be nearly equal to the threshold count, was way off, as much as 50% of the threshold count. After messing around for a while, the bottleneck ended up being the number of 'objects' being displayed on the worksheet. I'm using the term objects to refer to either a dot within a dot plot, a region, or simply a plot itself. The more of any of those object that the graphics card must process, the fewer the events it can process, therefore leading to a lot of data loss. It should follow then that if you display fewer objects, the data loss will decrease, and that's exactly what happened. We did one or two of the following to fix the problem: 1. Close all but a couple of dot plots in the worksheet to monitor integrity of data. 2. Change the "Events to Display" menu to a small number, e.g. 5000 events. 3. Do both. As soon as this was done, Processed events was right back to "normal" levels. This turns out to be a huge impact on overall yield for sorting as well because if it doesn't get processed, it doesn't get sorted. I suspect this is the reason why we've typically seen better yield on our MoFlo than we do on our Aria. I already knew that the sub par graphics card on these HPs caused the displaying of plots to lag after you collected your large files, but i didn't realize it was a bottleneck for data collection. Has anyone gone ahead and installed a high powered, really fast, graphics card in there standard HP to improve this issue? Can you send me info on what one(s) you've used? I'm using an HP XW4400s, and have noticed this on an Aria and LSRII. Do you think it's something other than the graphics processor? Thanks, Ryan ----------------------------------------------- Ryan Duggan Technical Director Flow Cytometry Facility University of Chicago Ph: 773.702.9212 or .5582 Web: http://ucflow.uchicago.edu Main Facility 910 E. 58th, Room 037 Chicago, IL 60637 ----------------------------------------------- This email is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this email message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is prohibited. If you have received this email in error, please notify the sender and destroy/delete all copies of the transmittal. Thank you.Received on Wed Apr 30 14:38:00 2008
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