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The Basics of Intracellular Cytokine Staining

Barry Moran

In this webinar you will learn:

  • The theory of intracellular cytokine staining.
  • Experimental design and sample preparation considerations.
  • Common pitfalls of intracellular cytokine staining.

Abstract:

Cytokine analysis via intracellular staining is arguably one of the most important technical advances in the immunologist’s toolbox in the last 20 years. It allows us to study the output of a number of different cytokines at a single cell level, at rates of thousands per second, which no other technique allows.

Generally this technique involves blocking the release of cytokines from the cell, labelling the cell with various surface markers to identify the cell type, and then fixing and permeabilising the cell to enable internalisation of the intracellular antibodies to bind with their corresponding cytokine.

Further, with the onset of more advanced cytometers, and the availability of a multitude of fluorescent molecules, we have the opportunity to look at hundreds of different cytokine combinations within a given population. However, with these advancements come more experimental and technical challenges.