FLOW CYTOMETER PERFORMANCE CHARACTERIZATION, STANDARDIZATION AND CONTROL

Published

May 9, 2017

Author(s)

Lili Wang, Robert A. Hoffman

Abstract

Flow cytometry is a widely used technique for the analysis of single cells and particles. It is an essential tool for immunological research, drug and device development, clinical trials, disease diagnosis, and therapy monitoring. However, the measurements made on different instrument platforms are often inconsistent resulting in unreliable measurements and impeding advances in biomedical research. This chapter showcases the methodologies to obtain key parameters for characterizing flow cytometer performance, sensitivity, background, electronic noise, and linearity. Further, various fluorescent beads, hard dyed and surface labeled are illustrated for the use of quality control, calibration, and standardization of flow cytometers. To compare instrument characteristics, fluorescence intensity units have to be standardized to MESF and ERF that are traceable to the existing primary fluorophore solution standards. With suitable biological controls or orthogonal method, users will be able to quantitatively measure DNA and RNA content per cell or biomarker expression in antibodies bound per cell. The comparable, reproducible and quantitative measurements using flow cytometers can only be accomplished upon instrument standardization through performance characterization and calibration, and use of proper biological controls.

Citation

Single Cell Analysis: Contemporary Research & Clinical Applications

Publisher Info

Springer-Nature, New York, NY

Pub Type

Books

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Keywords

Standard, calibrate, quality control, fluorescence, sensitivity, linearity, flow cytometer, MESF, ERF, ABC

Citation

Wang, L. and Hoffman, R. (2017), FLOW CYTOMETER PERFORMANCE CHARACTERIZATION, STANDARDIZATION AND CONTROL, Springer-Nature, New York, NY, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=922556 (Accessed December 26, 2024)