I forgot to comment on the issue of file format with the FC500. There is a problem with the file stucture of the FC500 files that we reported to Beckman-Couler a long time ago. They made an error in the classification of the parameter names in the FCS file structure. This does not have any effect if you analyze the data on their system, but as soon as you export their FCS histogram files to several other software packages (including Winmdi) ...its a problem becuase the parameter names are incorrect. (see below for a 2p histogram - both parmeter name data bit sizes are called $P2B - whereas the first one should be $P1B - so if you use another program to read these files, it may get confused by the double use of the $P2B...... further, there is no parameter in FCS 2.0 standard for word size with histograms - so this may cause some problems in defining the word size of the histogram files when you read them from 3rd party software programs....so if you use the fc500 listmode software to generate 2P histograms that are subsequently databased according to FCS nomenclature there could be a mix up od some names. Paul Robinson Purdue University -----------------------header of FC500 files------------------- FCS2.0 128 454 1024 9216 \$DATE\12-Mar- 04\$MODE\C\$BYTEORD\1,2\$NEXTDATA\0\$PAR\2\$TOT\ 81408\$DATATYPE\I\ $P1N\SS Log\$P1S\SS Log\$P1R\64\$P2B\16\$P2N\FS Log\$P2S\FS Log\$P2R\64\$P2B\16\$EXP\ \ $PROJ\ \$OP\gerald\$INST\ \$CYT\Cytomics FC 500\$SRC\ \$CELLS\ \@LOCATION\ \ $SYS\Cytomics(TM) RXP V1.0 for Win32\$BTIM\08:30:08\$ETIM\08:31:13\$RUNNUMBER\4137 \ ---------------------------------------------------------- On 2 Mar 2004 at 17:36, Stetler-Stevenson, Maryalice wrote: > Sorry I am late to reply on the problem with Coulter's FC500 files in FlowJo > not showing all parameters labeled, etc. The FC500 saves two copies of the > same data into one file. The first data set (FCS2) is the compensated "low > resolution" version. The second data set (FCS3) is the un-compensated "high > resolution" version. Part of the problem you described is that a number of > important keywords are missing for the "high resolution" version, making it > difficult to use other software. We saw this problem while doing an analysis > with WinList and notified Verity; they have added an option to address this > issue with the FC500 files. The data looks really good with the new version. > > > Maryalice Stetler-Stevenson > Chief, Flow Cytometry Unit > Laboratory of Pathology, NCI, NIH > > Sometimes you're the windshield, sometimes you're the bug. > J.Paul Robinson, PhD PH:(765)4940757 Professor of Immunopharmacology Professor of Biomedical Engineering Purdue University FAX:(765)4940517 EMAIL:jpr@flowcyt.cyto.purdue.edu WEB: http://www.cyto.purdue.edu Have you seen our new HCS webpage? http://www.cyto.purdue.edu/hcsReceived on Tue Mar 23 15:38:01 2004
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