Re: PerCP

Ray Hicks (rh208@cus.cam.ac.uk)
Fri, 10 May 1996 11:26:29 +0100

Hi Paul, We used cychrome extensively at my last lab, and it worked
fine, it has less spill over into FL1 than PE does, however fluorescein
goes into FL3 no matter what third reagent you use. Which way round
are you having your compensation problem? Due to B-D's compensation
circuitry, the only way to compensate FL1/FL3 is to compensate FL1/FL2
(both ways) then FL2/FL3 (both ways), which is straightforward enough
if you're using PE as well, but counter-intuitive if you're not. It
might be worthwhile getting your engineer to check that the machine is
aligned and fully functional if you can't get reasonable results, we
had problems doing three colour work with our FACSort a while ago (FL1
PMT 200 volts higher than FL2, FL2-%FL1 80% etc) and had to get have it
visited three or four times before the engineer agreed to replace our
FL1 PMT, after which our problems vanished.

Ray

At 4:55 pm 9/5/96, Paul J. Jansen wrote: >Paul J Jansen here, >
In the past we have used cychrome (??spell??) as a "third" >
color on our one laser Facscan. The problem we had with it was
>difficult to compensate between FL1 and FL3. We are considering using
>PerCP now and I would like to know if anyone has experience with it.
>BD has promised to throw in calibrite beads capable of doing the three
>color compensation. Comments? Questions? Suggestions? Thanx Paul J
>Jansen zinetti@acpub.duke.edu http://www.duke.edu/~zinetti DUMC
>Immunology http://www.duke.edu/~zinetti/Immunology/immuno1.html

Ray Hicks
________________________________________________________________________
|University of Cambridge |Tel 01223 330149
| |Department of Medicine |Fax 01223
336846 | |Level 5, Addenbrookes Hospital |e-mail
rh208@cus.cam.ac.uk | |Hills Road Cambridge |Web Page/
facsmac.med.cam.ac.uk | |CB2 |ftp
server 131.111.80.78 | |UK
| |
|_________________________________|_____________________________________|

Home Page Table of Contents Sponsors Web Sites
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