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Study of Cell-Matrix Adhesion Dynamics Using Surface Plasmon Resonance Imaging Ellipsometry

机译:表面等离子体共振成像椭圆光度法研究细胞-基质粘附动力学

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摘要

The interaction of cells with extracellular matrix, termed cell-matrix adhesions, importantly governs multiple cellular phenomena. Knowledge of the functional dynamics of cell-matrix adhesion could provide critical clues for understanding biological phenomena. We developed surface plasmon resonance imaging ellipsometry (SPRIE) to provide high contrast images of the cell-matrix interface in unlabeled living cells. To improve the contrast and sensitivity, the null-type imaging ellipsometry technique was integrated with an attenuated total reflection coupler. We verified that the imaged area of SPRIE was indeed a cell-matrix adhesion area by confocal microscopy imaging. Using SPRIE, we demonstrated that three different cell types exhibit distinct features of adhesion. SPRIE was applied to diverse biological systems, including during cell division, cell migration, and cell-cell communication. We imaged the cell-matrix anchorage of mitotic cells, providing the first label-free imaging of this interaction to our knowledge. We found that cell-cell communication can alter cell-matrix adhesion, possibly providing direct experimental evidence for cell-cell communication-mediated changes in cell adhesion. We also investigated shear-stress-induced adhesion dynamics in real time. Based on these data, we expect that SPRIE will be a useful methodology for studying the role of cell-matrix adhesion in important biological phenomena.
机译:细胞与细胞外基质的相互作用(称为细胞基质粘附)重要地控制着多种细胞现象。细胞基质粘附功能动力学的知识可以提供了解生物学现象的关键线索。我们开发了表面等离子体共振成像椭偏仪(SPRIE),以提供未标记活细胞中细胞-基质界面的高对比度图像。为了提高对比度和灵敏度,将空型成像椭圆仪技术与衰减全反射耦合器集成在一起。我们通过共聚焦显微镜成像证实了SPRIE的成像区域确实是细胞-基质粘附区域。使用SPRIE,我们证明了三种不同的细胞类型表现出不同的粘附特征。 SPRIE已应用于多种生物系统,包括细胞分裂,细胞迁移和细胞间通信。我们对有丝分裂细胞的细胞基质锚定进行了成像,从而为我们的知识提供了这种相互作用的第一个无标签成像。我们发现细胞间的通讯可以改变细胞基质的粘附,可能为细胞间的通讯介导的细胞粘附变化提供直接的实验证据。我们还实时研究了切应力引起的粘附动力学。基于这些数据,我们期望SPRIE将成为研究细胞基质粘附在重要生物学现象中的作用的有用方法。

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