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Experimental Techniques

Proliferation assay

The in vitro proliferation assay can be used to determine whether or not cells are triggered to divide after exposure to a specific stimulus, or to assess differences between cell populations in their ability to divide in response to the same stimulus.

Production of MHC Class I Tetramers

Major histocompatibility complex (MHC) tetramers were first described in the mid-1990s, and since then have been an invaluable tool to all T-cell immunologists. Identifying antigen-specific T cells is inherently difficult due to the low avidityof the interaction between T cell receptor (TCR) and MHC:peptide complex. The interaction has a half life of a mere 10 seconds!

Multiplex analysis of cytokines

The ELISA is a well-established method for quantifying a cytokine of interest in liquid samples. Multiplexing extends this to the measurement of multiple analytes in the same sample. Several multiplexing platforms exist, including bead-based (Luminex and flow cytometry cytokine bead array, CBA) and electrochemiluminescence (carbon surface, MSD) systems.

Flow Cytometry

Flow cytometry is a powerful tool to analyse multiple parameters on an individual cell basis. Cell populations can be characterised using a combination of antigens both on the surface and intracellularly. There are a number of practical applications regularly used by immunologists including immunophenotyping, measuring intracellular cytokine production, cellular proliferation, assessing cell viability and analysis of cell cycle, rare events, stem cells and fluorescent proteins. Cell sorting based on flow cytometry is used to separate cells into populations of interest.

Enzyme-linked immunosorbent assay (ELISA)

The enzyme-linked immunosorbent assay (ELISA) is an immunological assay commonly used to measure antibodies, antigens, proteins and glycoproteins in biological samples. Some examples include: diagnosis of HIV infection, pregnancy tests, and measurement of cytokines or soluble receptors in cell supernatant or serum. ELISA assays are generally carried out in 96 well plates, allowing multiple samples to be measured in a single experiment. These plates need to be special absorbant plates (e.g.