PMID: 374582Jan 1, 1979Paper

Fluorescence intensity resolution in flow systems

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
M J McCutcheon, R G Miller

Abstract

The factor which can limit fluorescence intensity resolution in a flow cytometer of the type in which cells pass perpendicularly through a focussed laser beam depends on signal intensity. For the brightest sources (e.g. fluorescent DNA stains), the coefficient of variation (CV) is limited in our system to around 3% by stream hydrodynamics, unstable illumination intensity, nonstoichiometric staining, etc. The weakest detectable sources (e.g. fluorescent cell-surface labels) are limited in coefficient of variation by shot noise in the photomultiplier due to constant background light levels. Finally, over a fairly wide brightness range between these extremes, resolution is determined primarily by photoelectron statistical variation on the signal itself (i.e. "photon statistics"). Thus photon collection and detection efficiency (solid angle, barrier filter passband, detector quantum efficiency) become of primary importance.

Citations

Jan 1, 1981·Journal of Immunological Methods·R G MillerG B Price
Mar 26, 2013·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·David NovoBartek Rajwa
Sep 1, 1980·Cytometry·J P Keene, B W Hodgson
Mar 1, 1985·Cytometry·P Ubezio, A Andreoni
Nov 1, 1980·Cytometry·S S StewartG B Price
Sep 15, 1982·Analytical Biochemistry·H S Kruth

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