Flow Cytometry

What is flow cytometry?

Last update: October 1st, 2019

Cytometry is a process in which physical and/or chemical characteristics of single cells, of other biological or nonbiological particles, are measured.

In flow cytometry, the measurements are made simultaneously as the cells or particles, suspended in fluid stream, pass individually through a beam of light of a measuring device, the flow cytometer. Some flow cytometers can physically separate cell subsets (sorting) based on their cytometric characteristics (cell sorters).

Flow cytometry can be used to count large numbers of cells or particles based on size, internal complexity, phenotype, cellular state, cell function, DNA content, gene expression, and to quantify these same cellular properties at a single-cell level. This allows to evaluate:

  1. Whether cells are present in a specimen
  2. How many are there
  3. What kinds of cells are they
  4. What their functional characteristics might be

OPTICAL MICROSCOPY

FLOW CYTOMETRY

Its working depends on the light scattering features of the cells under investigation, which may be derived from dyes or monoclonal antibodies targeting either extracellular molecules located on the surface or intracellular molecules inside the cell (see Flow Cytometry probes). This approach makes flow cytometry a powerful tool for detailed analysis of complex populations in a short period of time.

Resources

Publications:

  1. Adan A, et al. Flow cytometry: basic principles and applications. Crit Rev Biotechnol. 2017 Mar;37(2):163-176. Go to publication
  2. Cossarizza A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies. Eur J Immunol. 2017 Oct;47(10):1584-1797. Go to publication
  3. Piyasena ME, et al. Multinode acoustic focusing for parallel flow cytometry. Anal Chem. 2012 Feb; 84(4): 1831–1839. Go to publication
  4. Ward M, et al. Fundamentals of Acoustic Cytometry. Curr. Protoc. Cytom. 49:1.22.1-1.22.12. Go to publication
  5. Brown M and Wittwer C. Flow Cytometry: Principles and Clinical Applications in Hematology. Clinical Chemistry. 2000 April;46:8(B)1221–1229. Go to publication
  6. Howard M. Shapiro Practical flow cytometry, 4th ed. John Wiley hi Sons, Inc. 2003 August; 736 Pages. Go to publication