Slit scanning of Saccharomyces cerevisiae cells: quantification of asymmetric cell division and cell cycle progression in asynchronous culture.
Slit scanning flow cytometry has been applied to the analysis of the cell cycle and cell-cycle-dependent events in Saccharomyces cerevisiae, yielding information on the low-resolution spatial distribution of cellular components in single cells of unperturbed cell populations. Because this process is rapid, large numbers of cells can be analyzed to give distributions of parameters in a given population. To study asymmetric cell division and cell cycle progression, forward-angle light scattering (FALS) signals together with fluorescence signals from acriflavine-stained nuclei have been measured in cells from exponentially growing yeast populations. An algorithm has been developed that assigns the position of the bud neck in the FALS signals so that both FALS and DNA signals can be analyzed in terms of the contributions from the mother cell and the cell bud. The data indicate that mother cell FALS, on average, remains constant while FALS due to the cell bud increases as a cell progresses through the cell cycle. By identifying mitotic cells and measuring their properties, we have found that the coefficient of variation for the distribution of FALS is smallest within the dividing cell population and largest within the newborn cell population, in accordance with the critical size control mechanism of yeast cell growth. The use of this experimental approach to provide data for statistical population models is discussed.
Department of Chemical Engineering and Materials Science, University of Minnesota, St. Paul 55108.
D E
DE
Block
P D
PD
Eitzman
J D
JD
Wangensteen
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F
Srienc
D000167:Acriflavine; D000465:Algorithms; D002453:Cell Cycle; D002455:Cell Division; D002467:Cell Nucleus / Q000648:ultrastructure; D004271:DNA, Fungal / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D008027:Light; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics; D012542:Scattering, Radiation
D004271:DNA, Fungal; D005456:Fluorescent Dyes; D000167:Acriflavine