Investigating the Mechanics of Cell Division with A Side-View Atomic Force Microscope

用侧视原子力显微镜研究细胞分裂的机制

• 批准号: 1536736
• 负责人: Ovijit Chaudhuri
• 金额: $ 40万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1536736&HistoricalAwards=false
• 关键词: Investigating; Mechanics; Cell; Division; Side

Cell division is essential for life; it underlies the development of humans from embryo to full-grown adult, as well as regenerative processes such as wound healing and pathologies such as cancer. While cell division is an inherently mechanical process, its physical nature is poorly understood. This fundamental research will use a device called an "atomic force microscope" (AFM) to measure the small forces created by a cell and the stiffness changes of the cell during division into two cells. In addition, the AFM will be used to measure how the division of cells is changed by application of force. As cell division is an essential process in biology, the results of this work will contribute to understanding development and growth of organisms, tissue development, tissue regeneration, and cancer. The interdisciplinary approach taken in this research, involving the application of cutting edge techniques in engineering to the study of cell biology, will have a positive impact on the education of engineers and biologists including those from underrepresented groups. In addition, the novel measurement device built during the project will remain as a resource for future research by the current team and their colleagues.In tissues, cell division is physically restricted in all dimensions by neighboring cells and extracellular matrix. Dividing cells must be able to generate significant forces in a spatially and temporally coordinated manner to create the morphological changes accompanying cell division, and in some contexts must do this while experiencing physiological forces. How cells generate and respond to such forces remains unclear, as cell division has been primarily studied in cells cultured on flat petri dish surfaces. The objective of this project is to elucidate how cells generate and respond to forces, and alter their structural properties, during cell division. We will construct and use a high-resolution side-view imaging atomic force microscope to directly measure forces generated by cells during each stage of cell division while simultaneously imaging the dynamic cytoskeletal and chromosomal rearrangements in dividing cells in epi-fluorescence along a plane perpendicular to the sample surface. With this tool, we aim to address the following important questions regarding the mechanics of cell division: (1) What are the forces generated by cells following during cell division and how are they generated?; (2) How do the viscoelastic properties of cells change during each stage of cell division?; and finally (3) what is the effect of applied force on cell division?

细胞分裂对生命至关重要；它是从胚胎到成年成年人的人类发展的基础，以及再生过程，例如伤口愈合和癌症等病理。尽管细胞分裂是一种固有的机械过程，但其物理性质知之甚少。这项基本研究将使用称为“原子力显微镜”（AFM）的设备来测量细胞产生的小力以及分裂过程中细胞在两个细胞中的刚度变化。 此外，AFM将用于测量通过施加力改变细胞的分裂。由于细胞分裂是生物学的重要过程，这项工作的结果将有助于理解生物体的发育和生长，组织发育，组织再生和癌症。在这项研究中采用的跨学科方法涉及在工程研究中应用尖端技术在细胞生物学研究中的应用，将对工程师和生物学家的教育产生积极影响，包括来自代表性不足的群体的教育。 此外，在项目期间构建的新型测量设备将继续作为当前团队及其同事将来研究的资源。在组织中，细胞分裂在相邻细胞和细胞外基质中在所有维度上都受到物理限制。划分细胞必须能够以空间和时间协调的方式产生重要的力，以创建伴随细胞分裂的形态变化，在某些情况下，必须在经历生理力的同时这样做。细胞如何产生和对这种力的反应仍然不清楚，因为细胞分裂主要是在培养在培养皿表面上的细胞中的。该项目的目的是阐明细胞在细胞分裂期间的产生和响应和改变其结构特性的方式。我们将构建和使用高分辨率的侧视成像原子力显微镜直接测量细胞分裂每个阶段在细胞分裂的每个阶段产生的力，同时对沿垂直于样品表面的平面的Epi-Fluorescence中分裂细胞中的细胞中的动态细胞骨架和染色体重排。使用此工具，我们旨在解决有关细胞分裂力学的以下重要问题：（1）细胞分裂后细胞产生的力是什么？它们是如何产生的？ （2）在细胞分裂的每个阶段，细胞的粘弹性特性如何变化？最后（3）施加力对细胞分裂有什么影响？