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. 2001 Mar 27;98(8):4486–4491. doi: 10.1073/pnas.081075798

Figure 5.

Figure 5

Time-lapse microscopy of RK2-lacO and pUC-lacO segregation. Strains were grown in LB at 30°C, stained with FM 4–64 (red), and placed on an agarose slab; images were captured at various times by using a stage and microscope objective heated to 30°C as described in Materials and Methods. (Bars = 1 μm.) (A) Two time-lapse images taken 10 min apart of GFP-tagged RK2-lacO (JP872) showing the duplication of two foci into four foci (top cell) and the separation/migration of two midcell foci to the 1/4 and 3/4 cell positions (bottom cell). (B) Twenty images of JP872 (with GFP-tagged RK2-lacO) were collected 1 min apart; six of those images are shown. Three foci very close together at midcell at time 0 resolve into three clearly separate foci at midcell by the 7-min time point. Between the 9- and 10-min time points, the top two foci separate from the bottom focus more than 0.5 μm. Red shifting of GFP after repeated excitation yields foci that fluoresce both red and green, and therefore appear yellow when both colors are shown simultaneously. (C) Three time-lapse images of JP765 showing segregation of pUC-lacO. Two foci in one cell at time 0 migrate to the future midpoints of the two daughter cells by the 40-min time point.