Sheath buffers/H2O/PBS

Joseph Trotter - Salk Institute Flow Cytometry (trotter@flosun.sdsc.edu)
Tue, 16 Nov 93 10:05:27 PST

Flow people,

Be careful about the different refractive indices of sheath buffer
and sample buffer, you can introduce some nice "lens effects" at the
"wrong place" if the indices differ enough. It never hurts to match the
buffers fairly well, especially if scatter cv's are an issue. Running PBS
sample buffers in instruments using water as a sheath buffer sounds like a
formula for obtaining "less than optimal" cv's. We always switch out sheath
buffers to match the sample buffer refractive index if the sample buffer
isn't fairly "close" to our normal PBS buffer. Usually, this is only for
something like chromosomes or plant protoplasts, etc...

Since scatter measurements are largely a measurement of the difference
in refractive index between cells (or whatever) and the medium they are
suspended in, in makes no sense to surround the sample/buffer with a sheath
buffer having yet a different refractive index.

Joe Trotter
Salk Institute


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