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Curley bought several conjugates, and found that some of them, including
CD3 and CD45 were bright as can be. He couldn't figure out why conjugates of
CD33, CD10 and CD19 were so dim. He called Larry, who told him that his
problem was probably due to a bum PMT in FL3, after making sure that he was
using an appropriate filter, a 645 LPA. In fact, said Larry, I know of
several other instances where users had to struggle with their cytometer
manufacturers until they got a good PMT. One guy had to try 5 of them before
he found one that worked!
So Curley calls Moe, and tells him this, and Moe says bull#$%$! The
problem is the conjugate. It's going to be dimmer than what you're used to
seeing. For chrissakes, just look in their catalog...You excite the PE and
it emits at a peak of 575. You now ask it to excite the Cy5, which is
excited at 625-650. There ain't no way that you going to hit Cy5 with
anything that's going to excite it much. So, bag that junk and use something
that we make.
By now Curley's really flustered. You know, starts that banging himself on
the side of his head. But, since he's gotten a little smarter since his last
gig, and has discovered cyberspace, he decides to ask the rest of the
cytometry community for help.
With apologies to all the protagonists... any comments or suggestions would
be much appreciated.
--marc
Marc Langweiler 603-650-4924
DMS Pathology Lebanon, NH
langweiler@dartmouth.edu
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