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>Can anyone direct me to publications in which the time kinetics of
>binding of directly fluorochrome-conjugated antibody to living cells
>were followed by flow cytometry? The idea is to add the antibody to
>the cells and then immediately begin acquisition, without rinsing.
>Ideally, a single large file can be acquired over several minutes
>while binding proceeds; then medians can be taken on sequential
>"slices" of the data in the file. Thus, the entire kinetic curve is
>generated from a single sample.
>I'm particularly interested in the unrinsed mode described above, but
>I'd also be interested in any publications using a more conventional
>approach in which binding is stopped by rinsing at various times in
>different samples, and then each sample is analyzed as a single time
>point.
>Finally, I'd like to know what software is available (commercial or
>otherwise) to slice up the events in a single list-mode file. (I have
>some home-grown software under development if anyone is interested in
>using it.) Do any of the commercial packages (e.g. from Phoenix, Verity,
>B-D, or Coulter) do time-slicing?
The references below review the types of measurements Eric described,
give references to original work by Bohn & Mmanske, Crowell & Salzman,
Finney & Sklar and others, and give protocols for both equilibrium
methods and dilution methods. The KINPRO program in the Consort/VAX
package calculates gated means and standard deviations for time-slices
of FCS files and creates a file for plotting of these means versus time.
If CHRONOS is still supported for the HP, it does similar things.
R. F. Murphy (1990). Ligand Binding, Endocytosis, and Processing. In:
Flow Cytometry and Sorting, Second Edition, M. R. Melamed, T. Lindmo, M.
L. Mendelsohn (eds.), Wiley-Liss, Inc., New York, pp. 355-366.
R. F. Murphy (1992) Scatchard analysis by flow cytometry. In: Flow
Cytometry and Cell Sorting, A. Radbruch (ed.), Springer Verlag, Berlin,
pp. 59-62.
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