LCTM Image Gallery
This page contains an archive of all supplemental videos and corresponding papers.
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deRooij, J., A. Kerstens, G. Danuser, M. A. Schwartz and C. M. Waterman-Storer (2005) Integrin-dependent actomyosin contraction regulates epithelial cell scattering. J. Cell Biol. 171:153-164.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Contraction inhibitors destabilize cell-cell adhesion.
Video 2: GFP-E-cadherin during scattering.
Video 3: GFP-ZO-1 during scattering.
Video 4: Scattering on different types of ECM.
Video 5: Scattering does not depend on ß1 integrins.
Video 6: Substrate stiffness promotes scattering.
Wittmann, T. and C. M. Waterman-Storer (2005) Spatial regulation of CLASP affinity for microtubules by Rac1 and GSK3ß in migrating epithelial cells. J. Cell Biol. 169:929-939.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: 3xEGFP-CLASP2 in a PtK1 cell reveals MT plus end tracking in the cell body and dynamic lattice binding in the lamella.
Video 2: 3xEGFP-CLASP2 (green) and X-rhodamine-labeled tubulin (red) in a PtK1 cell demonstrating that all lamella MTs are CLASP decorated.
Video 3: Comparison of the dynamic behavior of EGFP-tagged CLASP2, EB1, CLIP-170, and APC in PtK1 cells.
Video 4: FRAP of EGFP-CLASP2 on lamella MT lattices (left) and growing plus ends in the cell body.
Video 5: 3xEGFP-CLASP2 in a PtK1 cell expressing constitutively active mRFP-Rac1(Q61L) showing randomized plus end tracking and lattice binding in both cell body and lamella.
Video 6: 3xEGFP-CLASP2 in a PtK1 cell treated with a Rac1 inhibitory peptide, TAT-Rac1(17-32), which inhibits MT lattice binding.
Video 7: 3xEGFP-CLASP2 in a PtK1 cell expressing constitutively active mRFP-GSK3ß(S9A).
Video 8: 3xEGFP-CLASP2 in a PtK1 cell in the presence of 20 mM lithium chloride, which induces ectopic MT lattice binding in the cell body.
Video 9: Truncated 3xEGFP-CLASP2(78-875) only tracks growing plus ends in PtK1 cells.
Torreano, P. A., C. M. Waterman-Storer, and C. S. Cohan (2005) The effects of collapsing factors on F-actin content and microtubule distribution of Helisoma growth cones.
Cell Motil. Cytoskeleton 60:166-179.
Video 1: Time-lapse movie (10 sec per frame, 10 min duration) of a growth cone labeled with x-rhodamine actin viewed with fluorescent speckle microscopy. ML-7 perfusion occurred during 2 out-of-focus frames (frames 19,20; 190 sec after start). First frame indicates location (*) of reorganized actin filaments apparent towards the end of the movie. After ML-7 application, note the progressive decrease in length and intensity of actin bundles. Also note that the undiminished rate of retrograde flow results in the movement and accumulation of orthogonal filaments around the C-domain.
Gupton, S. L., K. L. Anderson, T. P. Kole, R. S. Fischer, A. Ponti, S. E. Hitchcock-DeGregori, G. Danuser, V. M. Fowler, D. Wirtz, D. Hanein, and C. M. Waterman-Storer (2005) Cell migration without a lamellipodium: Translation of actin dynamics into cell movement mediated by tropomyosin.
J. Cell Biol. 168:619-631.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Cells containing skTM exhibit only one region of F-actin kinematic behavior at their leading edges.
Video 2: High levels of skTM inhibit the kinetic signature of the lamellipodium.
Video 3: High levels of skTM induce multiple filopodial protrusions from the cell edge.
Video 4: High levels of skTM induce changes in distribution and dynamics of paxillin-containing substrate adhesions.
Video 5: Cells with inhibited TM exhibit decreased lamellipodial protrusion persistence.
Yarar, D., C. M. Waterman-Storer and S. L. Schmid (2005) A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis.
Mol. Biol Cell 16:964-975.
Video 1: Time-lapse movie showing the formation and internalization of CCS (labeled with CLC-DsRed) imaged by TIR-FM (left panel) and WF-EFM (right panel). Scale bar is 1 µm. 2s/frame.
Video 2: Time-lapse movie showing the splitting of clathrin-coated vesicle from a larger CCS (labeled with CLC-DsRed) imaged by TIR-FM (left panel) and WF-EFM (right panel). Scale bar is 1 µm. 2s/frame.
Video 3: Time-lapse movie showing the s merging/coalescing of CCS with another CCS (labeled with CLC-DsRed) imaged by TIR-FM (left panel) and WF-EFM (right panel). Scale bar is 1 µm. 2s/frame.
Video 4: Time-lapse movie of CLC-DsRed (left panel) and Oregon Green actin (middle panel) and merged images [right panel, CLC (red) and actin (green)] during the formation and internalization of CCS. Both sequences were imaged by TIR-FM. Scale bar is 1 µm. 2.5s/frame.
Video 5: Time-lapse movie of CLC-DsRed (left panel) and Oregon Green actin (middle panel) and merged images [right panel, CLC (red) and actin (green)] during the internalization of a large CCS. Both sequences were imaged by TIR-FM. Scale bar is 1 µm. 2.5s/frame.
Video 6: Time-lapse movie of CLC-DsRed (left panel) and Oregon Green actin (middle panel) and merged images [right panel, CLC (red) and actin (green)] during the splitting and internalization of CCSs. Both sequences were imaged by TIR-FM. Scale bar is 1 µm. 2.5s/frame.
Video 7: Time-lapse movie of CLC-DsRed expressing Swiss 3T3 cells imaged by TIR-FM after 30 minute treatment with DMSO as a control (left panel), 5 µM latA (second from left) and then washed out (3rd from left), or 1 µM jasp (right panel). Scale bar is 5 µm. Time interval / frame is marked in minutes and seconds.
Video 8: Time-lapse movie of CLC-DsRed (left panel) and Oregon Green actin (middle panel) and merged images [right panel, CLC (red) and actin (green)] showing the recruitment of an actin-comet tail behind a motile CCS. Position of CCS in merged image is indicated by arrow. Scale bar is 1 µm. 2.5s/frame.
Ponti A., M. Machacek, S. L. Gupton, C. M. Waterman-Storer, and G. Danuser (2004) Two distinct actin networks drive the protrusion of migrating cells.
Science 305:1782-1786.
Video 1: Fluorescent Speckle Microscopy (FSM) time-lapse series of a newt lung epithelial cell migrating at the border of an epithelial monolayer. The cell was microinjected with a small amount of X-rhodamine labeled actin. Frame rate of raw movie: 10 sec/frame. Frame rate of replay: 5 frames/sec.
Video 2 : Raw FSM time-lapse series of a PtK1 cell migrating at the border of an epithelial monolayer. The cell was microinjected with a small amount of X-rhodamine labeled actin. Frame rate of raw movie: 10 sec/frame. Frame rate of replay: 5 frames/sec.
Video 3 : Animated map of F-actin retrograde flow in Video 1. A steady state representation of the same data is displayed Fig. 1A. Each frame of the animation is computed as the moving average of speckle trajectories over 5 frames (50 sec).
Video 4 : Animated map of F-actin retrograde flow in Video 2. A steady state representation of the same data is displayed in Fig. 1C. Each frame of the animation is computed as the moving average of speckle trajectories over 5 frames (50 sec).
Video 5 : Animated map of F-actin network turnover extracted from Video 1. A steady state representation of the same data is displayed in Fig. 1B. Each frame of the animation is computed as the moving average of the space-integrated kinetic scores associated with speckle appearances and disappearances (S6) over 5 frames (50 sec).
Video 6 : Animated map of F-actin network turnover extracted from Video 2. A steady state representation of the same data is displayed in Fig. 1D. Each frame of the animation is computed as the moving average of the space-integrated kinetic scores over 5 frames (50 sec).
Video 7 : Perfusion of PtK1 cells with 0.5 µM cytochalasin D. After disappearance of the lamellipodium, the F-actin at the leading edge retracts with the same velocity as the retrograde flow in the lamella. This suggests that low concentrations of cytochalasin D selectively remove the lamellipodium network, while the lamella network is more stable in the presence of inhibitors of polymerization.
Video 8 : Evolution of lamellipodium-to-lamella transition in Video 1. One of the frames is shown in Fig. 1E to illustrate the mathematical definition of the borderline based on profiles of F-actin velocity and turnover Figs 1F - G.
Video 9 : Evolution of lamellipodium-to-lamella transition in Video 2. Two of the frames are shown in Fig. 4B.
Vallotton P., C. M. Waterman-Storer, and G. Danuser (2004) Simultaneous mapping of filamentous actin flow and turnover in migrating cells by quantitative fluorescent speckle microscopy.
Proc. Natl. Acad. Sci. USA. 101:9660-9665..
Video 1: FSM raw images of a migrating newt lung epithelial cell filmed at 10-s intervals. Results of the analysis are shown in Fig. 1.
Video 2: Animation of the flow maps in steps of 30 s (compare Fig. 1B for a still version of the first map and Fig. 2 Upper for an enlargement of the pole region in three specific maps of the sequence).
Video 3: Animation of the turnover maps calculated according to Eq. 4 in steps of 30 s (compare Fig. 1D for a still version of the first maps and Fig. 2 Upper for an enlargement of the pole region in three specific maps of the sequence).
Video 4: Synthetic FSM images of a simulated meshwork contracting toward the center of the field of view. No meshwork turnover, leading to a rapid increase of fluorophore concentration in the polar region over time.
Video 5: Synthetic FSM images of a simulated meshwork contracting toward the center of the field of view. Meshwork depolymerization at a rate that increases toward to pole proportionally to the inverse of the to-pole distance. This rate conserves the density of filaments over time in presence of poleward flow.
Video 6: Synthetic FSM images of a simulated meshwork contracting toward the center of the field of view. Constant rate of meshwork depolymerization throughout the field of view, leading to a decrease of the average fluorophore concentration in the meshwork, yet a concentration increase in the polar region.
Video 7: Synthetic FSM images of a simulated meshwork contracting toward the center of the field of view. Constant rate of meshwork polymerization throughout the field of view, leading to an increase of the average fluorophore concentration in the meshwork.
Video 8: FSM raw images of a migrating newt lung epithelial cell treated with two perfusions of calyculin A filmed at 10-s intervals (compare the main text for the effect of the drug). Time points of drug application are indicated by title frames.
Video 9: Animation of flow velocity (vectors) and flow speed (colored background) maps calculated for selected time points of the original frame sequence. Time points of drug application are indicated by title frames.
Video 10: Animation of the turnover maps calculated for selected time points of the original frame sequence. Time points of drug application are indicated by title frames.
Schlunck, G., H. Damke, W. B. Kiosses, N. Rusk, M. H. Symons, C. M. Waterman-Storer, S. L. Schmid and M. A. Schwartz (2004) Modulation of Rac function by dynamin.
Mol. Biol. Cell. 15:256-267.
Video 1: Figure 5 A
Video 2: Figure 5 E
Video 3: Figure 7 A
Video 4: Figure 7 C
Prigozhina, N. and C. M. Waterman-Storer (2004) Protein kinase D-mediated anterograde membrane trafficking is required for fibroblast motility.
Curr. Biol. 14:88-98.
© Elsevier Science Ltd. All rights reserved.
Video 1: Dynamics of PKD-kd-GFP-labeled membrane tubules emanating from the TGN region in a living Swiss 3T3 cell.
Video 2: Anterograde transport of VSVG-GFP in a control Swiss 3T3 cell.
Video 3: VSVG-GFP dynamics in a Swiss 3T3 cell coexpressing GST-tagged PKD-kd.
Video 4: Blockage of anterograde secretion pathway by PKD-kd-GFP expression inhibits cell motility.
Wittmann, T., G. M. Bokoch and C. M. Waterman-Storer (2004) Regulation of microtubule destabilizing activity of Op18/stathmin downstream of Rac1.
J. Biol. Chem. 279:6196-6203.
Video 1: Comparison of microtubule dynamics in vitro in the presence of 1.5 µM Op18/stathmin (left) or 2.0 µM Pak1-phosphorylated Op18/stathmin (right).
Vallotton,
P., A. Ponti, C. M. Waterman-Storer, E. D. Salmon and G. Danuser (2003) Recovery, visualization, and analysis of actin and tubulin polymer flow in live cells: A fluorescent speckle microscopy study.
Biophys. J. 85:1289-1306.
Video 1: F-actin speckle flow in a migrating newt lung epithelial cell (left: original FSM movie, right: residual movie after subtraction of tracked speckles).
Video 2: Microtubule speckle flow in a Xenopus egg extract mitotic spindle (left: original FSM movie, right: residual movie after subtraction of tracked speckles).
Wittmann, T., G. M. Bokoch and C. M. Waterman-Storer (2003) Regulation of leading edge microtubule and actin dynamics downstream of Rac1.
J. Cell Biol. 161:845-851.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Microtubules in a control PtK1 cell.
Video 2: Microtubules in a constitutively active Rac1(Q61L)-expressing PtK1 cell.
Video 3: Microtubules in a dominant negative Rac1(T17N)-expressing PtK1 cell.
Video 4: Direct comparison of microtubule dynamics in PtK1 cells expressing Rac1(Q61L) or Rac1(T17N).
Video 5: Microtubule fragments exhibiting net treadmilling in a Rac1(Q61L)-expressing PtK1 cell.
Video 6: Microtubules in a PtK1 cell injected with the Pak inhibitory fragment PBD/ID(H83L).
Video 7: Microtubules in a PBD/ID(H83L)-injected, Rac1(Q61L)-expressing PtK1 cell.
Video 8: Actin dynamics in a Rac1(Q61L)-expressing PtK1 cell.
Video 9: Actin dynamics in a PBD/ID(H83L)-injected, Rac1(Q61L)-expressing PtK1 cell.
Salmon, W. C., M. C. Adams and C. M. Waterman-Storer (2002) Dual-wavelength fluorescent speckle microscopy reveals coupling of microtubule and actin movements in migrating cells.
J. Cell Biol. 158:31-37.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Correlation of time-lapse actin FSM to fixed phalloidin staining.
Video 2: Time-lapse FSM reveals four distinct zones of polymerization and movement behavior of f-actin.
Video 3: Dual-wavelength FSM of microtubules (green) and f-actin (red).
Video 4: Retrograde flow of microtubules oriented perpendicular to the leading edge in the lamellipodium is coupled to the movement of immediately adjacent lamellum speckles.
Video 5: Retrograde flow of microtubules oriented perpendicular to the leading edge in the lamellipodium is not coupled to the movements of immediately adjacent lamellipodium f-actin.
Video 6: Microtubules grow along f-actin bundles toward dense f-actin plaques.
Video 7: A microtubule with a quiescent-end move in association with a moving f-actin bundle seen as a linear array of f-actin.
Zhou, F. Q., C. M. Waterman-Storer and C. S. Cohan (2002) Focal loss of actin bundles causes microtubule redistribution and growth cone turning.
J. Cell Biol.157:839-849.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Microtubule dynamics in a polylysine-attached Helisoma growth cone.
Waterman-Storer, C. M., D. Y. Duey, K. L. Weber, J. Keech, R. E. Cheney, E. D. Salmon and W. M. Bement (2000) Microtubules remodel actomyosin networks in Xenopus egg extracts via two mechanisms of F-actin transport.
J. Cell Biol. 150:361-376.
This research was originally published in The Journal of Cell Biology. © The Rockefeller University Press.
Video 1: Alignment of f-actin networks in Xenopus extracts containing nocodazole.
Video 2: F-actin zippering alignment in Xenopus extracts containing nocodazole.
Video 3 does not exist.
Video 4 : Astral microtubule ejection and plus end-directed gliding.
Video 5: Expansion of a sperm aster by microtubule ejection and gliding.
Video 6: F-actin network deformation in the presence of randomly oriented microtubules.
Video 7: Clearing of f-actin from around a sperm aster.
Video 8: Microtubule-dependent F-actin jerking motility.
Video 9: Jerking motility of f-actin moving in association with a gliding microtubule.
Video 10: Microtubule/Actin interactions in Xenopus egg extracts, microtubules nucleated from a sperm aster.
Video 11: Sperm aster microtubules and f-actin in the presence of anti-cytoplasmic dynein antibodies.
Video 12: Straight gliding of f-actin along the microtubule lattice.
Video 13: Pure microtubules moving on a kinesin-coated coverslip in the presence of a network of f-actin bundled by alpha-actinin.
Waterman-Storer, C. M., W. C. Salmon and E. D. Salmon (2000) Feedback interactions between cell-cell adherens junctions and cytoskeletal dynamics in newt lung epithelial cells.
Mol. Biol. Cell. 11:2471-2483.
Video 1: Microtubule dynamics are inhibited in contacted newt lung epithelial cells.
Video 2: Photoactivation of caged fluorescein tubulin in a contacted newt lung epithelial cell.
Video 3: Comparison of f-actin dynamics in contacted and migrating newt lung epithelial cells using Fluorescent Speckle Microscopy (FSM).
Waterman-Storer, C. M. (1998) Microtubules and Microscopes: How discoveries about microtubule dynamics in living cells have been led by the development of imaging technologies.
Mol. Biol. Cell. 9:3263-3271.
Video 1: Time-lapse fluorescence microscopy of rhodamine microtubule dynamics in a PtK1 cell recorded with an ISIT camera.
Video 2: VE-DIC of the lamella of a newt lung epithelial cell.
Video 3: Photoactivation of C2CF-tubulin fluorescence in the mitotic spindle of a PtK2 cell.
Video 4: Microtubule dynamics in a Xenopus neuronal growth cone recorded under anoxic conditions with a cooled CCD camera.
Video 5: Dual fluorescence digital imaging of the dynamics of microtubules and endoplasmic reticulum in the lamella of a newt lung epithelial cell.
Video 6: DE-DIC and fluorescence overlaid images of microtubule and cell surface dynamics in a newt lung epithelial cell.
Video 7: Microtubule fluorescent speckle imaging of the lamella of a newt lung epithelial cell.
Waterman-Storer, C. M., Desai, A., J. C. Bulinski and E. D. Salmon (1998) Fluorescent speckle microscopy, a method to visualize the dynamics of protein assemblies in living cells.
Curr. Biol. 8:1227-1230.
© Current Biology Ltd.
Video 1: Fluorescent speckle imaging: Actin in the lamella of a migrating epithelial cell.
Waterman-Storer, C. M., and E. D. Salmon (1998) Endoplasmic reticulum membrane tubules are distributed in living cells by three distinct microtubule dependent mechanisms.
Curr. Biol. 8:798-806.
© Current Biology Ltd.
Video 1: Microtubule-Endoplasmic reticulum interactions in a migrating newt lung epithelail cell.
