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Each bead trajectory was also modeled with the simplest viscoelastic model (Fig. 2) which realistically described the cytoplasmic environment during creep (F≠0), as described in ref. [28].
The magnetic force, which was constant along the entire bead trajectory within a given experiment, ranged from 22 to 280 pN.
Each bead trajectory was approximated as a sum of an exponential and a linear function of time: x t) = A·(1−e−B·t)+C·t, where x is bead displacement, t is time and A, B and C are fitting parameters.
The bead trajectory was recorded with a CoolSNAPfx video camera (Photometrics, Tucson, AZ) controlled by Image Pro Express Media CyberneticsSan Diego, CA Cand and subsequently tracked with in-house developed particle-tracking software.
In seven of eight control KVs, beads followed regular, anti-clockwise paths whereas bead trajectory was qualitatively abnormal in 6 of 10 ofd1 MO-injected vesicles, deviating from a circular anti-clockwise path with loops and zig-zags (Fig. 5D and E and Supplementary Movies 1 and 2).
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Figures 1a, 1b show examples of the bead trajectories before adding drug.
It follows that the removal of all out-of-plane bead trajectories from the top layer data has a significant impact on the average MSD, G′ and G″ of the top layer (Fig. 3(a)).
In order to improve the depth resolution of the images, we remove bead trajectories from the uniform data, which experience a drop in intensity above 20%, as determined in the previous section.
The reason for this is that out of the bead trajectories removed from the bottom layer data (44 beads removed from a total of 185), it's unlikely that many of the removed beads are from the top layer of cells due to the asymmetry in bead visibility (Fig. 2).
During acquisition, the bead followed a trajectory determined by the lateral drift of the setup.
For empirically tracing each constituent particle trajectory, spot spraying bead was produced.
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