From: Wayne Shreffler (wayne.shreffler@mssm.edu)
Date: Mon Dec 22 2003 - 17:57:08 EST
I think I have experienced the problem you are describing with 32-bit data files from Diva. The data are uncompensated whether you use the Diva compensation matrix or not, but I have experienced this 'overcompensation' artifact though I have not thought that it might only occur when using the acquisition compensation matrix. I have spoken with Tree Star and they were quite responsive, but I think they were unable to replicate the problem. We never came to a solution, and I have since found a work around. In my experience, this overcompensation occurs when files are re-loaded into the memory cache and the compensation is recalculated. By changing the default prefs to compensate only when needed, increasing the memory allocation for the cache, and working with small groups of files to prevent , I have largely avoided the problem -- though it is a bit inconvenient. Wayne G. Shreffler, MD, PhD Assistant Professor Department of Pediatrics Division of Allergy & Immunology Mount Sinai Medical Center, box 1198 One Gustave L Levy Place New York, NY 10029 212-241-7244 voice 212-426-1902 fax On Friday, December 19, 2003, at 03:01 PM, Andy Johnson wrote: > Hello > > Can I just ask if anybody else is having trouble converting DiVa files > into similar FlowJo files ? > > There appears to be no problem when files are not compensated in DiVa. > The problem occurs when DiVa compensated files are read in FlowJo > resulting in "massive" over compensation of the data. Unfortunately we > have got little help from FlowJo other than try their new > version...which we have with similar results. > > We are looking at 8 colours plus, so is it just that the compensation > matrix of the DiVa and flowJo clash ? > > We could save uncompensated data in DiVa and look at that in > FlowJo...but for the price you pay, this should not have to be the > case ? Plus we are saving large files of rare events, so this would > remove a large portion of our sorting sample. > > Any suggestions ? > > Andy > Merry Christmas > -- > Andy Johnson > FACS Facility Manager > Biomedical Research Centre > UBC > Vancouver > 604-822-7838 >
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