Investigation of Cellular Compliance Sensing and Response Using Single-Cell-Pico-Force-Microscopy

这项工作将促进我们对细胞如何处理机械信号的理解。这项工作是基于单细胞微力显微镜(SCPFM)，这是一种有望广泛应用于许多细胞生物学研究的新仪器。SCPFM将纳米机电系统(NEMS)的近单分子测力能力与集成的微流控技术相结合，实现了对单个细胞的精确流体控制，从而实现精确的药物刺激。了解细胞力的产生和调节对于了解细胞功能的许多基本方面至关重要；其中包括：细胞运动、细胞分裂和细胞组织；生物体功能的许多方面：伤口愈合、炎症和胚胎发生；以及许多疾病，包括许多癌症和心血管疾病。具体地说，这项工作将研究细胞外基质(ECM)的顺应性感知和力响应，作为研究力学在细胞调节中的作用的模型系统。例如，SCPFM能够详细和系统地研究单个片状脂体对药理和机械刺激的分子力学反应。在这项工作中，将获得顺应性感知和响应事件的力时间记录，并将这些记录用于组装分子机械力签名库，用于分析力时间记录。SCPFM将器件物理、纳米制造和电气工程与表面化学、细胞生物学和生物化学结合在一个复杂的、相互关联的系统中。这为研究生和博士后提供了一个非同寻常的机会，使他们成为能够领导和成功整合跨学科团队和项目的跨学科科学家。本科生和少数民族学生将有机会通过加州理工大学S冲浪和MURF项目(暑期/少数民族本科生研究奖学金)参与其中。这项研究的影响将由参与加州理工大学S课堂连接的研究生和博士后带到当地社区，后者将研究人员与当地高中科学课堂合作，在那里他们作为大学预科学生的榜样。

This work will advance our understanding of how cells process of mechanical signals. The effort is based upon Single-Cell-Pico-Force-Microscopy (SCPFM), a new instrument poised to become widely applicable to many studies in cell biology. SCPFM integrates near-single-molecule force measurement capabilities of Nano Electro Mechanical Systems (NEMS) with integrated microfluidics enabling precision fluid control, and thereby precision pharmacological stimulation, of individual cells. Understanding cellular force generation and regulation is critical to understanding many basic aspects of cell function; among these are: cell motility, cell division, and cell organization; many aspects of organism function: wound healing, inflammation, and embryogenesis; and numerous diseases, including many cancers and cardiovascular disease. Specifically, this work will investigate the compliance sensing and force response of the Extra-Cellular-Matrix (ECM) as a model system for studying the role of mechanics in cell regulation. SCPFM enables, for example, detailed and systematic study of the molecular-mechanical responses of individual lamellipodia to pharmacological and mechanical stimulation. Force-time records of compliance sensing and response events will be acquired in this work, and these will be used to assemble a library of molecular-mechanical force signatures with which to analyze the force-time records. SCPFM combines device physics, nano-fabrication, and electrical engineering with surface chemistry, cell biology and biochemistry in a complex, interconnected system. This presents an extraordinary opportunity for graduate students and post-docs to become interdisciplinary scientists capable of leading and successfully integrating interdisciplinary teams and projects. Undergraduate and minority students will have the opportunity to be involved through Caltech?s SURF and MURF programs (Summer/Minority Undergraduate Research Fellowship). The impact of this research will be carried into the local community by the graduate students and post-docs involved in cooperation with Caltech?s Classroom Connection, which partners researchers with local high school science classes where they serve as role models for pre-college students.