Cellular functions are what cells do to stay alive, proliferate, or defend themselves. Before the advent of flow cytometry, functional assays were done on multiple platforms and were labor intensive. In the last several years, flow cytometry-based functional assays have emerged as fast, precise and customizable assays that can be used in basic, preclinical or clinical research settings.
Check out these examples of flow cytometry-based functional assays that are currently in use across multiple therapeutic research areas.
Proliferation
Cell proliferation assays use a cell-permeable fluorescent dye that is taken up by parent cells and diluted in daughter cells during subsequent cell divisions. Flow cytometry analysis reveals different peaks of fluorescence that correspond with each round of cell division, and this data can be used to assess the proliferative capacity of different cell populations.
Endocytosis, Exocytosis, and Phagocytosis
Cells take in and release molecules from the cell surface through different cellular transport mechanisms that use subcellular structures such as vesicles. Endocytosis is the process by which cells take up molecules and complexes at the plasma membrane into vesicles that are trafficked into the cytoplasm. Phagocytosis is a form of endocytosis by which extracellular pathogens can be taken into cells and destroyed. Exocytosis is the trafficking of vesicles from inside the cell to the plasma membrane, resulting in the fusion of membranes and the release of the vesicle contents to the extracellular domain. Each of these mechanisms can be measured by different flow cytometry assays that use fluorescently labeled molecules like dextran or fluorescent bacteria.
Calcium Flux
Intracellular calcium levels in resting cells are typically stable, and cellular stimuli trigger the flux of intracellular calcium ions. Specialized dyes that bind to free calcium ions can be used to measure changes in intracellular calcium and give insight into the activation state of cells.
Oxidative Metabolism
Normal physiological processes cause oxidative metabolism and produce different byproducts, including oxygen and peroxide. Cancer and inflammation can cause a dramatic increase in oxidative metabolism, and flow cytometry can be used to measure these byproducts through the use of fluorescent cell permeable probes that bind specifically to different oxygen species.
Considering adding these types of functional assays to your next research project to gain new insights by using a trustworthy and robust flow cytometry-based research platform.
