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Intracellular cytokine detection by fluorescence-activated flow cytometry: basic principles and recent advances

Giulia Freer1, Laura Rindi

  • 1Department of Experimental Pathology, University of Pisa, Via San Zeno, I-56127 Pisa, Italy. giulia.freer@med.unipi.it

Methods (San Diego, Calif.)
|April 16, 2013
PubMed

Insights

Intracellular cytokine staining is a flow cytometry method to analyze T cell function and phenotype. This technique is vital for vaccine trials and infectious disease research, offering detailed cellular insights.

Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Intracellular cytokine staining is a flow cytometry technique.
  • It involves culturing stimulated cells with a protein secretion inhibitor, followed by fixation, permeabilization, and fluorescent antibody staining.
  • Modern flow cytometers can detect up to 18 colors simultaneously.

Purpose of the Study:

  • To review the features of intracellular cytokine staining.
  • To provide a standard laboratory protocol for this technique.
  • To discuss recent applications in immunological studies.

Main Methods:

  • Stimulation of cytokine-producing cells.
  • Inhibition of protein secretion.
  • Fixation and permeabilization of cells.
  • Staining with fluorescent antibodies for intracellular cytokines and cell markers.

Main Results:

  • Enables simultaneous determination of antigen-specific T cell function and phenotype.
  • Allows measurement of cell proliferation and viability.
  • Highly popular method for assessing antigenicity in vaccine trials and infectious diseases.

Conclusions:

  • Intracellular cytokine staining is a versatile and powerful immunological technique.
  • It is a key method for understanding immune responses.
  • Its applications continue to expand in various research fields.

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