Re: using flow for B cell/plasma cell evaluation

Simon Monard (monard@phri.nyu.edu)
Tue, 2 Apr 1996 11:56:38 -0500

Hi Jason

I can't quite understand what you want to do. B-Cells which encounter an
antigen which binds their antigen receptor (surface Ig) will produce a
calcium flux which can be measured by flow providing there are enough
B-cells with that antigen receptor. We have demonstrated that using HEL
specific B-Cells from transgenic mice. The frequency of primary B-Cells
specific to a single antigen would be too low to measure calcium fluxes
unless you could pre-enrich for them. We prepared cells from mouse spleens
stained all the cells we didnt want, gated them out and looked at the
calcium signal from the rest. Some of this work is published in Science vol
271 pp348-340. Humans however are often uncooperative about giving up their
spleens.

Good luck

Simon

Simon Monard
FACS Lab Manager
Aaron Diamond Center
455 1st Avenue
New York
NY 10016

ph (212) 340 3741

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