Tracking of individual cell cohorts in asynchronous Saccharomyces cerevisiae populations.
A novel flow cytometric procedure has been developed with the aim to obtain the growth properties of individual Saccharomyces cerevisiae cells in asynchronous culture. The method is based on labeling of the cell surface with FITC-conjugated concanavalin A and detection of the single-cell fluorescence with flow cytometry after cell exposure to growth conditions. Because the formation of new cell wall material in budded cells is restricted to the bud tip, exposure of the stained cells to growth conditions results in three cell types: (i) stained cells, (ii) partially stained cells, and (iii) unstained cells. Analysis of the staining pattern over time permits the determination of the specific growth rate of the cell population, the length of the budded cell cycle phase, and the growth pattern during the cell cycle of newly formed, partially stained daughter cells. The procedure has been tested with yeast cell populations growing at different rates. The data suggest an exponential increase in the size of individual cells during the cell cycle, as reflected by the forward angle light scattering (FALS) signals. It has been found that the apparent single-cell specific cell size growth rates, determined by FALS intensity, are significantly lower than the specific growth rates of the overall population. This could indicate that the tracking of a cohort of cells is significantly perturbed by a distribution of staining levels of daughter cells at cell division and that FALS may not be a good indicator of the cell size.
Porro
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Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis/St. Paul 55108, USA.
Srienc
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D002453:Cell Cycle; D002455:Cell Division / Q000502:physiology; D002462:Cell Membrane / Q000737:chemistry; D002469:Cell Separation; D003208:Concanavalin A; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000166:cytology*
C042784:fluorescein isothiocyanate-concanavalin A; D003208:Concanavalin A; D016650:Fluorescein-5-isothiocyanate