Flow Cytometry

Flow cytometry is a technology commonly used in biomedical research to measure physical and chemical properties of single cells. Cells in suspension are stained with various fluorescent dyes that bind to a specific macromolecule and pass through a laser beam one by one. Next, the equipment measures scattered and emitted light intensity that can be translated to size, granularity, expression levels of protein, RNA, or DNA content. In this chapter, we explain the principles of flow cytometry and provide a basic protocol to measure cell surface protein expression levels with fluorescent antibodies. As a practical example, we demonstrate how flow cytometry can be used to detect changes of cellular populations during the process of induced pluripotent cell generation.