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Fig. 1. Microtubule arrays in the fission yeast cell cycle are depicted by cartoon, still images and kymograph. (A) Description of how to construct a kymograph from a series of time-lapse images using DeltaVision software: 1. A series of time-lapse images were taken of a cell of interest. 2. The images were rotated to orient the microtubule of interest along the horizontal axis. 3. The images were then cropped around the cell boundary and the time-lapse images were converted into a 3D stack. 4. The stack of images were rotated 90° around the horizontal axis to align the time points in a vertical stack; this allows for the changes in microtubule length overtime to be seen as a line trace or kymograph. 5. The kymograph was then rotated so that time is on the horizontal axis. (B) The top panel shows a schematic demonstrating the different microtubule arrays as they appear in various stages of the cell cycle. The middle panel shows still images of a cell expressing GFP:
-tubulin (YY105) taken from a time-lapse movie of a cell progressing from G2 through telophase. Images correspond to the cartoon drawings and to the areas marked on the kymograph (lower panel): (a) G2-interphase, (b) G2-M transition with stage 1 spindle, (c) phase 2 spindle, (d) early anaphase B, (e) late anaphase B, and (f) telophase. Bar, 2 µm. The lower panel shows a kymograph made from the same time-lapse movie as the images shown in the middle panel. The images a-f in the middle panel correspond to the indicated regions of the kymograph. The kymograph is presented with time in minutes progressing along the horizontal axis and the length of the cell on the vertical axis. The three spindle phases are labeled below the kymograph. The transition from G2-M is marked with an arrow. The white outline marks triangular area of fluorescence that marks the life history of an interphase half bundle. Bar, 2 µm.
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