What is d in visualisation of collective cell mobilisation by the white wave' method?
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Fig. 1. Visualisation of collective cell mobilisation by the 'white wave' method. (A)In vitro scratch-wound healing assay using mIMCD3 cells. Representative images taken at the indicated time points post wounding (p.w.) are displayed. (B)Scheme of the 'white wave' method. Cartoons of a sheet of cells moving into the denuded region (gray) (a), and the same cell sheet a short time interval later (b). The first four rows of cells from the wound move forward, whereas the next two rows remain stationary. We calculate the absolute value of the difference between a and b by subtracting individual pixel differences, and display the result in a new 'difference' image (c). (C)Application of the 'white wave' method to an image of an mIMCD3 cell sheet 1 hour p.w. and one image from 15 minutes later. Bracket shows the white area in which cells are moving within this 15 minute interval. (D)Mobilisation of the cells in the experiment shown in A at the indicated time points, visualised using the 'white wave' method. The 'difference image at time T' is derived by subtracting the phase-contrast image at time T from that at T+T (here, T15 minutes-this is true for all the other data, unless otherwise stated). Dashed lines show the propagation of the 'white wave' of cell mobilisation; bracket shows the 'black area' where cells are starting to halt at 18 hours p.w. The bare substrate seen in the denuded area appears black, because it remains unchanged during the experiments. However, occasionally some white pixels appear because of cell debris from the wound drifting in the medium. (E)Schematic of d and. For details, see the Results section and supplementary material Fig. S2. To avoid the complexities associated with the immediate white difference readout (D), measurement of commences 1 hour after wounding for all our experiments. (F)Representative difference images of E1 cells at the indicated time points after wounding. Here, T45 minutes (see also supplementary material Fig. S1). Note that the 'white wave' does not propagate back into this 'fibroblastlike' cell sheet. Scale bars: 200m (A,D), 100m (C,F)
Fig. 1. Visualisation of collective cell mobilisation by the 'white wave' method. (A)In vitro scratch-wound healing assay using mIMCD3 cells. Representative images taken at the indicated time points post wounding (p.w.) are displayed. (B)Scheme of the 'white wave' method. Cartoons of a sheet of cells moving into the denuded region (gray) (a), and the same cell sheet a short time interval later (b). The first four rows of cells from the wound move forward, whereas the next two rows remain stationary. We calculate the absolute value of the difference between a and b by subtracting individual pixel differences, and display the result in a new 'difference' image (c). (C)Application of the 'white wave' method to an image of an mIMCD3 cell sheet 1 hour p.w. and one image from 15 minutes later. Bracket shows the white area in which cells are moving within this 15 minute interval. (D)Mobilisation of the cells in the experiment shown in A at the indicated time points, visualised using the 'white wave' method. The 'difference image at time T' is derived by subtracting the phase-contrast image at time T from that at T+T (here, T15 minutes-this is true for all the other data, unless otherwise stated). Dashed lines show the propagation of the 'white wave' of cell mobilisation; bracket shows the 'black area' where cells are starting to halt at 18 hours p.w. The bare substrate seen in the denuded area appears black, because it remains unchanged during the experiments. However, occasionally some white pixels appear because of cell debris from the wound drifting in the medium. (E)Schematic of d and. For details, see the Results section and supplementary material Fig. S2. To avoid the complexities associated with the immediate white difference readout (D), measurement of commences 1 hour after wounding for all our experiments. (F)Representative difference images of E1 cells at the indicated time points after wounding. Here, T45 minutes (see also supplementary material Fig. S1). Note that the 'white wave' does not propagate back into this 'fibroblastlike' cell sheet. Scale bars: 200m (A,D), 100m (C,F)
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