Picturing single T cells and its cytokine production
The assay is performed in a multitestplate, in which the membrane in each well is coated with specific antibodies against the cytokines to detect.
To activate the T cells, antigen or mitogen solution is mixed in each well with the isolated PBMCs.
The antigen-specific, activated T cells produce cytokines which are bound by the coated antibodies.
The secrected cytokines are detected by a secondary, conjugated antibody and a following chromogen.
The resulting spots in the wells are corresponding to the number of antigen-specific T cells.