Figure 1.
Budding yeast cell progression through the cell cycle.
Figure 2.
Evolution of cell density and identification of growth stages before DNA staining.
Five growth stages were defined according to log OD of the cell culture: 1: lag phase (black), 2: start of growth (cyan), 3: exponential stage (green), 4: slowdown phase (orange), 5: stationary phase (red).
Figure 3.
Time courses of the relative proportions of the cell populations during the various cell cycle stages, using SYTOX® Green fluorochrome (G0: black; G1: purple; S: green and G2/M: magenta).
Colored scale indicates the stages as defined in Figure 2.
Figure 4.
Evolution of the cytometric signal during cell culture (time in min) with A: cell size (Forward SCatter), B: SYTOX® Green, C: propidium iodide (PI), D: TO-PRO-3, E: 7-aminoactinomycin D (7-AAD) and F: SYBR® Green I.
The x axes are in arbitrary units. The y axes correspond to relative frequencies adjusted for each signal. Each histogram represents about 20000 cells Colors correspond to the different growth stages as defined in Figure 2. For B to F, the vertical lines indicates the median signal (coefficient of variation) corresponding strictly to yeast cells at G1 stage during the exponential phase.
Figure 5.
One dimensional representation of the apparent cell size (A, B), and DNA content as revealed by SYTOX Green (C, D) and SYBR Green I (E, F) labeling.
Arrows underline the main evolutions of the DNA content from the lag phase to exponential growth (A, C, E) and from exponential growth to the stationary phase (B, D, F). Colors correspond to the different stages as defined in Figure 2.
Table 1.
Signal intensity with different fluorochromes during the exponential cell growth phase (G1 and G2/M) or stationary phase (G0).
Figure 6.
Two dimensional representation (DNA staining signal/apparent size) used to determine gating parameters with different fluorochromes (SYTOX® Green, PI, TO-PRO®-3, 7-AAD and SYBR® Green I).
Gating was optimized for the whole growth experiment, and is summarized by the representation of yeast populations at three typical stages: exponential (245 min), slowdown (485 min) and stationary (725 min