UNIT 1.3 Standardization, Calibration, and Control in Flow Cytometry

  1. Lili Wang1,
  2. Robert A. Hoffman2

Published Online: 5 JAN 2017

DOI: 10.1002/cpcy.14

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Wang, L. and Hoffman, R.A. 2017. Standardization, calibration, and control in flow cytometry. Curr. Protoc. Cytom. 79:1.3.1-1.3.27. doi: 10.1002/cpcy.14

Author Information

  1. 1

    Biosystems and Biomaterials Division, NIST, Gaithersburg, Maryland

  2. 2

    Independent Consultant, Livermore, California

Publication History

  1. Published Online: 5 JAN 2017


Because flow cytometers are designed to measure particle characteristics, particles are the most common materials used to calibrate, control, and standardize the instruments. Definitions and cautions are provided for common terms to alert the reader to critical distinctions in meaning. This unit presents extensive background on particle types and cautions and describes practical aspects of methods to standardize and calibrate instruments. Procedures are provided to characterize performance in terms of optical alignment, fluorescence and light scatter resolution, and sensitivity. Finally, suggestions follow for analyzing particles used for calibration. © 2017 by John Wiley & Sons, Inc.


  • standardization;
  • calibration;
  • control;
  • particles;
  • MESF (molecules of equivalent soluble fluorochrome);
  • ERF (equivalent number of reference fluorophores);
  • sensitivity;
  • resolution