re: analysis for dem fluorescence
Gero Brockhoff t866623 (Gero.Brockhoff@klinik.uni-regensburg.de)
Fri, 3 Mar 1995 13:21:07 +0100
Dear Susan,
in previous studies we have determined % positive cells with fairly dim fluorescence compared to an isotype control.
Our measurements were performed on the FACScan (Beckton Dickinson) and analysis was done with the Lysis II software (also Beckton Dickinson) by normalized subtraction described in detail as follows:
. First you have to determine the exact number of interested cells in seperated histograms by histogram statistics (after adequate gating of debris and dubletts).
. The next step is to overlay the histgrams of interest (first the staining, second the isotype control)
. Then you can adjust the number of events with multiply by in the histogramm tools menue.
. After doing this, you have to subtract in the histogramm tools menue.
. Remaining cells are concerned as % positive of your measurement.
I hope this helps,
with best regards
Gero Brockhoff
e.mail: brckhoff@klinik.uni-regensburg.de
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
