tetramer assay on XL or ELITE

From: Kristi Harkins (kharkins@iastate.edu)
Date: Tue Aug 24 1999 - 17:55:52 EST


Hi from Iowa,

  I am looking for published references and anecdotal information on the use
of a Beckman Coulter XL or ELITE instrument in rare event detection.  We
have a client that would like to perform a tetramer assay of MHC/peptide
multimers bound to CD3/CD8 positive T-cells.  As discussed in a previous
thread this assay  is a rare event detection of from 20 in 200K to 2 in 200K
cells based upon detection of bound fluorescent multimers.  The references
I've found utilized a FACS Caliber for this assay.  

Questions include: 
How do you setup and verify the instrument for accuracy in detection?
What is a reasonable total cell count to analyze for statistical accuracy? 
Is available disk space the only limitation to the size of a listmode file
accumulated on the XL?
What are some good references for rare event detection on the XL or ELITE?

Thanks in advance and I'll summarize the replies if enough respond. - Kristi
Kristi R. Harkins, PhD
Director, Cell & Hybridoma Facility
Iowa State University
1104 Molecular Biology Building
Ames, IA 50011
Office (515) 294-2472
Lab    (515) 294-8504
FAX    (515) 294-0453
http://www.biotech.iastate.edu/facilities/cellhyb/homepage.htm



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