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

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.