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Fig. 3. Spatiotemporal mapping of pseudopod formation. (A) Sequence of fluorescent images of GFP-ABD-expressing, vegetative wild-type cells recorded at intervals of 250 ms and shown every 15 seconds. Bar, 10 µm. (B-E) Cell boundary activity of the cell in A was mapped in polar coordinates (left-right corresponding to 0-360 degrees) along the time axis (from top to bottom over 135 seconds). (B) Membrane velocity, (C) F-actin intensity and (D) radial distance from the centroid. (E) The same images as B-D, coloured in red, green, and blue, respectively were merged. Areas of the protruding sites are shown at higher magnification in supplementary material Fig. S2. To improve appearance of the structures, the original sequence was low-pass filtered at 1 Hz using a windowed-sinc filter (see http://rsb.info.nih.gov/ij/plugins/windowed-sinc-filter.html). Bleb formation was detected as bright areas in B and dark areas in C at the timing of emergence (resulting in red parts in E) as shown with the arrowheads in B, C and supplementary material Fig. S2. The arrows in A indicate persisting filopodia that also appear in D.
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