Lattice diffusion of CLIP-170(H2)–GFP protein (a carboxyl truncation of the plus end microtubule tracking protein CLIP-170). The movie shows pseudocolored overlaid images of rhodamine-labeled microtubules (red) and GFP-tagged CLIP-170(H2) protein (green). In the absence of EB1, CLIP-170(H2) binds along the length of the microtubule lattice and exhibits diffusive movement (blue arrowhead). (Scale bar, 2 micron.) Microtubule dynamics were visualized at 22 °C using a TIRF microscope ﬁtted on an inverted microscope (Nikon TE-2000U). The laser light intensities used for imaging were kept low, so that the photobleaching rates were slow compared with the dwell times. Images were captured with a backilluminated electron-multiplying CCD camera (Cascade-512B, Photometrics) using time-lapse capture at burst mode at 10 frames per second for the analysis of +TIP dynamics at the single-molecule resolution. Movie corresponds to video S2 from Proc Natl Acad Sci U S A. 2009 Jan 13;106(2):492-7. Epub 2009 Jan 6.
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