The enzyme-linked immunospot (ELISPOT) assay is a flexible and highly sensitive tool for analyzing the immunological secretions of peripheral blood and lymphoid cell populations. This process allows for the detection, enumeration, and characterization of individual cytokine-secreting cells within cultured cell populations.
In the case of a cytokine assay, a culture plate is coated with anti-cytokine antibodies. The cells are introduced, under conditions which cause them to secrete the cytokines. These cytokines bind to the antibodies in question, after which the cells are removed. An enzyme-labeled anti-cytokine is then applied to the bound cytokines, and a chromogenic (i.e. color-generating) substrate is applied to anti-cytokines. This substrate creates colored spots which can be detected via machine vision. (This process is graphically depicted here.)
In order to be truly effective though, the software techniques used must be designed to make maximal use of machine vision techniques and man-machine interface usability. This can be tricky, as the individuals who develop the software are generally not the types who would be using it on a regular basis. UI design is an art form unto itself, and its difficulty is compounded when such disparate fields and biochemistry and software engineering are involved.
This necessitates the development of readers
or analyzers which can perform the analysis at high speed, and with high accuracy. This avoids most of the tedium involved in this process, and if scientifically validated statistical techniques are used, the subjectivity involved can be eliminated as well.
Information on reagent sets and an expanding glossary of immunological terms might prove useful to the layman.