The key to these products is to remember your ratios.
You know how many beads are in the tube.
You know what volume of cells you put into the tube.
After that, it doesn't matter what you dilute the tube up to, as long as you
don't do any washing steps. The ratio of cells to beads will remain
constant.
If you do washing steps, you will lose cells and beads.
I'd recommend using Coulter's Flow-Count beads. For one, they're a lot
cheaper, and two, you can add them after your washing steps. If you have to
wash, you have to wash. There's nothing you can do about it. Might as well
get beads that work better for what you're doing.
http://www.beckmancoulter.com/Immunotech/Systems-b6.asp?spec=Fluorospheres&N
ame=Flow%2DCount
Good luck
Kb
--
Science is built with facts as a house is with stones--but a collection of
facts is no more a science than a heap of stones is a house. -Jules Henry
Poincare (1854-1912)
Keith Bahjat
Graduate Assistant
University of Florida
College of Medicine
Gainesville, Florida
Phone: (352) 392-4887
Fax: (352) 392-5393
kbahjat@ufl.edu
Hi List,
I have a question about Becton Dickinson's Trucount product. The tubes
come with no product insert so I went on their Web page
http://www.bdfacs.com/source_book/html/23_3025.shtml. I found a procedure
for absolute CD4 counts which I have tried to adapt to a cell death
assay.When calculating absolute counts, their procedure says use a test
volume which is found on the foil package. There is no test volume listed,
just the number of beads per lot. Has anyone else used this product to
determine absolute counts for tissue culture with reproducible results?
After getting weird results we contacted BD who states that the
procedure is wrong so I guess you cannot count on the accuracy of their Web
site. I'm pretty bitter after spending lots of time and money on this.
Paula
Paula Fukushima
Flow Cytometry
LP, NCI, NIH
10 Center Drive MSC-1500
Bethesda, Md. 20892-1500
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:57:43 EST