Mandal Kalpana, Asnacios Atef, Goud Bruno, Manneville Jean-Baptiste
Institut Curie, Paris Sciences et Lettres Research University, CNRS, UMR 144, F-75005 Paris, France.
Sorbonne Universités, Université Pierre et Marie Curie, University Paris 06, CNRS, UMR 144, F-75005 Paris, France.
Proc Natl Acad Sci U S A. 2016 Nov 15;113(46):E7159-E7168. doi: 10.1073/pnas.1605112113. Epub 2016 Oct 31.
The mechanical properties of cells impact on their architecture, their migration, intracellular trafficking, and many other cellular functions and have been shown to be modified during cancer progression. We have developed an approach to map the intracellular mechanical properties of living cells by combining micropatterning and optical tweezers-based active microrheology. We optically trap micrometer-sized beads internalized in cells plated on crossbow-shaped adhesive micropatterns and track their displacement following a step displacement of the cell. The local intracellular complex shear modulus is measured from the relaxation of the bead position assuming that the intracellular microenvironment of the bead obeys power-law rheology. We also analyze the data with a standard viscoelastic model and compare with the power-law approach. We show that the shear modulus decreases from the cell center to the periphery and from the cell rear to the front along the polarity axis of the micropattern. We use a variety of inhibitors to quantify the spatial contribution of the cytoskeleton, intracellular membranes, and ATP-dependent active forces to intracellular mechanics and apply our technique to differentiate normal and cancer cells.
细胞的力学特性会影响其结构、迁移、细胞内运输以及许多其他细胞功能,并且已证实在癌症进展过程中这些特性会发生改变。我们开发了一种方法,通过结合微图案化和基于光镊的主动微流变学来绘制活细胞的细胞内力学特性。我们用光镊捕获内化在铺有弩形粘性微图案的细胞中的微米级珠子,并在细胞发生阶跃位移后跟踪它们的位移。假设珠子的细胞内微环境服从幂律流变学,从珠子位置的弛豫来测量局部细胞内复剪切模量。我们还用标准粘弹性模型分析数据,并与幂律方法进行比较。我们发现,沿着微图案的极性轴,剪切模量从细胞中心到周边以及从细胞后部到前部逐渐降低。我们使用多种抑制剂来量化细胞骨架、细胞内膜和ATP依赖性主动力对细胞内力学的空间贡献,并应用我们的技术来区分正常细胞和癌细胞。