A novel cell force sensor for quantification of traction during cell spreading and contact guidance
A novel cell force sensor for quantification of traction during cell spreading and contact guidance
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DOI：
10.1529/biophysj.106.093302
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发表时间：
2007-07-01
影响因子：
3.4
通讯作者：
Gold, J.
中科院分区：
文献类型：
--
作者：
Tymchenko, N.;Wallentin, J.;Gold, J.
在这项工作中，我们提出了一个脊，微加工，力传感器，可用于研究细胞之间的机械相互作用表现出接触指导和底层细胞培养基质，和力传感器性能的功能验证评估。该基板包含阵列的垂直支柱之间的固体脊，微制造的硅晶片使用光刻和深反应离子蚀刻。通过原子力显微镜测量柱的弹簧常数。对于延时实验，将细胞接种在柱撑基底上，并在配备有re.反射微分干涉对比光学系统。内皮细胞（EC）和。在附着、铺展和迁移过程中观察到成纤维细胞。开发定制图像分析软件以解析细胞边界、细胞与柱的对齐和迁移、单个柱的位移，并量化细胞牵引力。联系指导分类。阳离子基于细胞相对于微加工脊的排列和移动角度以及细胞伸长。在与脊细胞力传感器的初步调查，我们已经观察到接触指导EC，但不是在。成纤维细胞在接触引导和非接触引导但移动的EC之间观察到机械力的最大幅度的差异。然而，需要进一步的实验来确定这一观察结果的统计学意义。偶然间，我们观察到细胞行为的另一个特征，即细胞力方向的逆转。四舍五入下测得的力的方向。成纤维细胞从早期细胞贴壁时的向外变化到进一步观察伸展期时的向内变化。下测量的力的范围。broblasts（高达138 nN）大于在EC中测量的（高达57 nN），表明刚性硅传感器能够分辨大范围的力，因此能够检测细胞类型之间的牵引力的差异。这些观察结果表明脊状细胞力传感器诱导接触引导的功能证明，并且在刚性硅中构建的柱撑细胞力传感器具有必要的灵敏度来检测不同细胞表型和形态之间的牵引力矢量的差异。
In this work, we present a ridged, microfabricated, force sensor that can be used to investigate mechanical interactions between cells exhibiting contact guidance and the underlying cell culture substrate, and a proof-of-function evaluation of the force sensor performance. The substrates contain arrays of vertical pillars between solid ridges that were microfabricated in silicon wafers using photolithography and deep reactive ion etching. The spring constant of the pillars was measured by atomic force microscopy. For time-lapse experiments, cells were seeded on the pillared substrates and cultured in an on-stage incubator on a microscope equipped with re. flected differential interference contrast optics. Endothelial cells (ECs) and. broblasts were observed during attachment, spreading, and migration. Custom image analysis software was developed to resolve cell borders, cell alignment to the pillars and migration, displacements of individual pillars, and to quantify cell traction forces. Contact guidance classifi. cation was based on cell alignment and movement angles with respect to microfabricated ridges, as well as cell elongation. In initial investigations made with the ridged cell force sensor, we have observed contact guidance in ECs but not in. broblast cells. A difference in maximal amplitude of mechanical forces was observed between a contact-guided and non-contact-guided, but mobile, EC. However, further experiments are required to determine the statistical significance of this observation. By chance, we observed another feature of cell behavior, namely a reversion of cell force direction. The direction of forces measured under rounded. broblast cells changed from outwards during early cell attachment to inwards during further observation of the spreading phase. The range of forces measured under. broblasts (up to 138 nN) was greater than that measured in EC (up to 57 nN), showing that the rigid silicon sensor is capable of resolving a large range of forces, and hence detection of differences in traction forces between cell types. These observations indicate proof-of-function of the ridged cell force sensor to induce contact guidance, and that the pillared cell force sensor constructed in rigid silicon has the necessary sensitivity to detect differences in traction force vectors between different cell phenotypes and morphologies.