Many in vitro assays have been developed to screen and evaluate the efficacy of immune cell-mediated killing. The most common of these is the release assay where effector cell-mediated disruption of the target cell membrane results in leakage of its cytoplasmic contents into the culture medium. Endogenous biomolecules (such as lactate dehydrogenase) or previously added exogenous labels (such as the radioisotopes 51Cr or 111In) that leak into the media are then measured as an indirect readout of the damage caused by effector cells.
Alternative endpoint methods include flow cytometry, ELISA-based granzyme measurement, and morphometric analyses by microscopy. Although traditional end-point assays still serve as important tools for evaluating the efficacy of immune cell-mediated killing, there is a critical need for more efficient, homogeneous, automated assays (requiring less hands-on time) that aren’t hampered by the artifacts and hassle of using labels and that can provide quantitative results.
ACEA’s xCELLigence Real-Time Cell Analysis (RTCA) instruments utilize gold microelectrodes embedded in the bottom of microtiter wells to non-invasively monitor cell status including cell number, shape/size and attachment. The major distinguishing features of this technology include enhanced sensitivity, the preclusion of labels and, importantly, kinetic measurement of cell health/behavior.
Read the application note.