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BP: Fachverband Biologische Physik
BP 22: Physics of Cells I
BP 22.5: Vortrag
Mittwoch, 16. März 2011, 16:30–16:45, ZEU 250
High-Precision Dynamics of Membrane Protrusions and Dorsal Ruffles in Mouse Fibroblasts — •Erik Bernitt1, Pritpal Singh2, Christina Oettmeier1, Cheng-Gee Koh2, and Hans-Günther Döbereiner1 — 1Institut für Biophysik, Universität Bremen, Germany — 2School of Biological Sciences, Nanyang Technological University, Singapore
High-contrast microscopy techniques like total internal reflection fluorescence microscopy allow to accurately locate cellular structures. Cell dynamics can thus be precisely described using advanced localization algorithms in combination with an appropriate tracking method. We implemented a novel velocity chart method and applied it to spreading NIH 3T3 fibroblast cells. We could clearly identify a difference in cell spreading velocities of wildtype cells and cells that overexpress the PAK phosphatase POPX2. The precision of the method is thereby only limited by spatial and temporal resolution of the micrographs and thus superior to the traditional method of kymographs that relies on a preserved direction of structure propagation. We give a detailed analysis of experimental error in measuring membrane protrusion speeds. Further, we report on our latest advances in the quantification of the dynamics of dorsal ruffles that are a characteristic feature of POPX2-overexpressing cells. Function and mechanism of dorsal ruffles are still under discussion and quantitative dynamic data is missing. Therefore our data provides the basis for the establishment of models describing dorsal ruffle dynamics.