The main in vitro methods for immunologists to analyze cells of the immune system are flow cytometry, PCR and various forms of ELISA (e.g. ELISPOT). Together, these techniques allow measurement of different cell populations at the molecular and functional level (“immuno-phenotyping”) and quantification of immune responses, for example, cytokine release. While these platforms are extremely powerful, they provide little insight into morphology and the spatial interactions of cells. Moreover, as ‘endpoint’ measures, they are not well suited to report on changes in biology over time.
Over recent years, real-time live-cell analysis has become an enabling research method for immunologists, addressing many of the limitations described above. Here, we review 10 functional bio-assay “applications” that illustrate the breadth, depth and value of this approach.