This chapter discusses the cell cycle analysis by flow cytometry. Growth of cell populations can be analyzed by considering individual cells to progress sequentially through a series of compartments or phases. The processes of mitotic division and DNA synthesis can be recognized by light microscopy and by incorporation of [3H] thymidine followed by autoradiography, respectively. Application of these methods led to a description of the cell cycle composed of four phases: mitosis (M), gap 1 (G1) preceding DNA synthesis, synthesis of DNA (S), and gap 2 (G2) following DNA synthesis and preceding the next mitosis. The chapter reviews flow cytometric principles, and then describe selected methods for the preparation of cultured cells for flow cytometry. Three examples of the use of flow cytometry in cell cycle studies are also given. The method of autoradiography and the analysis of labeling index and mitotic index experiments are described. A brief discussion of flow cytometric concepts, including cell transport, fluorescence excitation and detection, cell sorting, and data aquisition, emphasizing features that are important in DNA-content measurements, is presented.
ASJC Scopus subject areas
- Molecular Biology