MEMS device for applying shear and tension to an epithelium combined with fluorescent live cell imaging
MEMS device for applying shear and tension to an epithelium combined with fluorescent live cell imaging
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DOI：
10.1088/1361-6439/abb12c
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发表时间：
2020-12-01
影响因子：
2.3
通讯作者：
Pruitt，Beth L.
中科院分区：
文献类型：
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作者：
Garcia，Miguel A.;Sadeghipour，Ehsan;Pruitt，Beth L.
机械力在细胞和组织的生物功能中起着重要作用。虽然许多研究已经探测了细胞的力响应并测量了细胞产生的力，但它们主要集中在拉伸力，而不是剪切力。在这里，我们描述的设计，制造和应用的硅微机械设备，能够独立地施加和感应在上皮细胞单层的张力和剪切力。我们将该设备与立式显微镜集成，以在机械扰动后的数小时内实现活细胞明场和细胞荧光成像。使用增加刚度和相同的位移输入的设备，我们表明，上皮细胞表现出伴随更高的最大阻力张力和更快的力松弛。此外，我们还表征了上皮对循环剪切载荷的力响应。虽然循环剪切扰动下的上皮细胞的最大阻力在循环之间保持不变，但循环载荷导致阻力更快地松弛。这里提出的装置可以应用于研究其他单层形成细胞类型的力响应，并且与细胞结构和功能的药理学扰动兼容。
Mechanical forces play important roles in the biological function of cells and tissues. While numerous studies have probed the force response of cells and measured cell-generated forces, they have primarily focused on tensile, but not shear forces. Here, we describe the design, fabrication, and application of a silicon micromachined device that is capable of independently applying and sensing both tensile and shear forces in an epithelial cell monolayer. We integrated the device with an upright microscope to enable live cell brightfield and fluorescent imaging of cells over many hours following mechanical perturbation. Using devices of increasing stiffness and the same displacement input, we demonstrate that epithelia exhibit concomitant higher maximum resistive tensile forces and quicker force relaxation. In addition, we characterized the force response of the epithelium to cyclic shear loading. While the maximum resistive forces of epithelia under cyclic shear perturbation remained unchanged between cycles, cyclic loading led to faster relaxation of the resistive forces. The device presented here can be applied to studying the force response of other monolayer-forming cell types and is compatible with pharmacological perturbation of cell structures and functions.