Biophysics Institute, University of Bremen, Otto-Hahn Allee, Bremen, Germany.
Biophys J. 2011 Mar 16;100(6):1420-7. doi: 10.1016/j.bpj.2011.01.063.
We report on the characterization of actin driven lamellipodial protrusion forces and velocities in keratocytes. A vertically mounted glass fiber acted as a flexible barrier positioned in front of migrating keratocytes with parallel phase contrast microscopy. A laser beam was coupled into the fiber and allowed detecting the position of the fiber by a segmented photodiode. Calibration of the fiber was carried out with the thermal oscillation method. Deflection and force signals were measured during lamellipodial protrusion. Velocity was constant during initial contact whereas loading force increased until finally the cell was stalled at higher forces. Stall forces were on the order of 2.9 ± 0.6 nN, which corresponds to a stall pressure of 2.7 ± 1.6 nN/μm(2). Assuming a density of actin filaments of 240 filaments per μm, we can estimate a stall force per actin filament of 1.7 ± 0.8 pN. To check for adaption of the cell against an external force, we let the cell push toward the glass fiber several times. On the timescale of the experiment (∼1 min), however, the cell did not adapt to previous loading events.
我们报告了在角膜细胞中肌动蛋白驱动的片状伪足伸出力和速度的特性。一根垂直安装的玻璃纤维充当了一个灵活的障碍物,放置在正在迁移的角膜细胞前面,并用平行相差显微镜进行观察。激光束被耦合到纤维中,并允许通过分段光电二极管检测纤维的位置。通过热振荡法对纤维进行校准。在片状伪足伸出期间测量挠度和力信号。在初始接触期间速度保持恒定,而加载力增加,直到细胞最终在较高的力下停止。停止力约为 2.9 ± 0.6 nN,相当于 2.7 ± 1.6 nN/μm(2)的停止压力。假设肌动蛋白丝的密度为每 μm 240 根丝,我们可以估计每根肌动蛋白丝的停止力为 1.7 ± 0.8 pN。为了检查细胞对外部力的适应能力,我们让细胞多次推向玻璃纤维。然而,在实验的时间尺度(约 1 分钟)内,细胞并没有适应之前的加载事件。