Rapid, Simultaneous Measurement of Dna, Protein, and Cell Volume in Single Cells from Large Mammalian Cell Populations

The advent of high speed, cell analysis systems (1-5) for rapid measurement of physical and biochemical properties in single cells has provided new and useful techniques for performing a wide variety of biological experiments . Such systems recently have been used in studies related to DNA content per cell as a function of chromosome number (6, 7), effects of chemotherapeutic agents on cell cycle traverse (8), and quantitation of cell surface binding of the plant lectin concanavalin A (9) as well as immunofluorescent detection of antigen-binding cells (10) . These investigations have involved quantitative fluorescent staining of monodisperse cell populations in liquid suspension by techniques specific for biochemical components of interest and subsequent rapid analysis of the fluorescence emission signal obtained as each cell traverses a laser beam . General operating characteristics of this flow system, as well as validation of the methodology, have been presented in detail elsewhere (7, 11) . These earlier studies involved measurement of only a single cellular property. In this report we describe a method for measurement and simultaneous analysis of DNA, protein, and cell volume in the same cell by use of a newly developed multiparameter cell analysis system which incorporates several pre-existing analytical techniques and allows all the various measurements to be performed on the same cell under conditions where large populations can be analyzed rapidly and a high degree of statistical precision obtained . Electronic processing of fluorescence and volume signals from each cell provides a direct method for establishing relationships between various cellular properties reflecting specific phases of the cell cycle .