Flow cytometry is well suited to DNA analysis because dyes are available that bind DNA in a proportional and linear fashion. This allows the quantitation of DNA content, enabling the identification of normal diploid cells at rest, those that are actively synthesizing DNA, and those that are either premitotic or actually in mitosis. Once these phases of the cell cycle have been identified, antibody detection of cell cycle-related proteins can be combined with DNA content determination to relate protein expression with stages of the cell cycle. Addition of the thymidine analog, bromodeoxyuridine, during cell growth will also allow the identification of cells that are actively synthesizing DNA. Chromosome numbers, whether aberrant or not, can also be determined using DNA content analyses. Thus, both ploidy (e.g., in megakaryocytes) or aneuploidy (e.g., in neoplastic disease) can be assessed. Protocols, with their advantages and limitations, are described here by.
Key Words: Aneuploidy; cell cycle; checkpoints; mitosis; ploidy.
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