Innovative Tools for Three Dimensional Traction Force Microscopy of Single Cells

单细胞三维牵引力显微镜的创新工具

• 批准号: 9465846
• 负责人: Ottmar Klaas
• 金额: $ 14.94万
• 依托单位: SIMMETRIX, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-21 至 2019-03-20
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9465846
• 关键词: Accounting; Algorithms; Alpha Cell; Atherosclerosis; Atomic Force Microscopy; Behavior; Benchmarking; Biochemical; Biological; Biology; Biomechanics; Breast Cancer Cell; Cell Shape; Cell physiology; Cells; Chronic; Code; Communities; Computer Systems; Computer software; Computer-Aided Design; Custom; Data Set; Development; Dimensions; Disease Progression; Elements; Embryonic Development; Event; Extracellular Matrix; Fibroblasts; Gel; Generations; Heterogeneity; Human; Individual; Lead; Link; Malignant Neoplasms; Measurable; Measures; Mechanics; Methods; Microspheres; Modeling; Pathway interactions; Performance; Physics; Physiological; Play; Population; Process; Property; Research; Resolution; Role; Signal Transduction; Software Design; Spatial Distribution; Specific qualifier value; Techniques; Tissues; Traction; Variant; biological systems; cancer cell; cell motility; cell type; computer program; design; improved; innovation; insight; novel; novel therapeutic intervention; optical imaging; response; software development; stem cell differentiation; tool; Accounting; Algorithms; Alpha Cell; Atherosclerosis; Atomic Force Microscopy; Behavior; Benchmarking; Biochemical; Biological; Biology; Biomechanics; Breast Cancer Cell; Cell Shape; Cell physiology; Cells; Chronic; Code; Communities; Computer Systems; Computer software; Computer-Aided Design; Custom; Data Set; Development; Dimensions; Disease Progression; Elements; Embryonic Development; Event; Extracellular Matrix; Fibroblasts; Gel; Generations; Heterogeneity; Human; Individual; Lead; Link; Malignant Neoplasms; Measurable; Measures; Mechanics; Methods; Microspheres; Modeling; Pathway interactions; Performance; Physics; Physiological; Play; Population; Process; Property; Research; Resolution; Role; Signal Transduction; Software Design; Spatial Distribution; Specific qualifier value; Techniques; Tissues; Traction; Variant; biological systems; cancer cell; cell motility; cell type; computer program; design; improved; innovation; insight; novel; novel therapeutic intervention; optical imaging; response; software development; stem cell differentiation; tool

Project Summary Tractions exerted by individual cells on their surroundings play a critical role in mechanical events in biology such as tissue contraction, folding, cell shape changes, or cell movements, and in many basic cellular functions such as biochemical signaling, proliferation, and differentiation. These processes are in turn implicated in the progression of diseases like cancer, atherosclerosis, and other chronic fibrotic conditions. Recently, this remarkable link has been utilized to develop exciting new therapeutic interventions that rely on disrupting mechano-signaling machinery within the cell, and the pathways that lead to the remodeling of the extra-cellular matrix (ECM). Techniques that can precisely quantify the spatial variation and heterogeneity of cellular traction within and between cells have found important applications in understanding and controlling these processes. Of these, three-dimensional traction force microscopy (3D TFM) has emerged as a particularly valuable tool since it is applied to cells embedded in a three-dimensional ECM, the natural state for most cells. Current 3D TFM approaches are challenged by the critical steps of using optical images to generate a 3D geometrical model of the matrix surrounding the cell, and inferring cellular tractions from displacement estimates of micro-beads embedded in the matrix. Approximations incurred in these steps lead to significant errors in computed tractions that in turn lead to erroneous biological conclusions. Thus there is critical need to develop more accurate and high resolution 3D TFM techniques. The long-term objective of the proposed research is to improve and automate the 3D TFM process so that it can be effectively used to answer mechanobiological questions and design new therapeutic interventions. This will be accomplished by (a) applying advanced segmentation and mesh generation techniques to optical images to generate 3D geometric models and finite element meshes of the matrix surrounding a cell, and (b) by developing and implementing new algorithms to determine the spatial distribution of cellular tractions from measured micro-beads displacements, while accounting the nonlinear elastic response of the matrix. These developments will be validated through benchmark studies, and their utility will be demonstrated by quantifying the traction exerted by cancer cells embedded in a synthetic extracellular matrix.

项目摘要 单个细胞在周围环境中施加的文章在生物学的机械事件中起着至关重要的作用 例如组织收缩，折叠，细胞形状变化或细胞运动，以及在许多基本的细胞中 诸如生化信号传导，增殖和分化之类的功能。这些过程又是 与癌症，动脉粥样硬化和其他慢性纤维化疾病等疾病的进展有关。 最近，这种非凡的链接已被用来开发依赖的令人兴奋的新治疗干预措施 破坏细胞内的机械信号机械，以及导致重塑的途径 细胞外基质（ECM）。 可以精确量化细胞牵引力的空间变化和异质性的技术 在细胞之间发现了在理解和控制这些过程中的重要应用。其中， 三维牵引力显微镜（3D TFM）已成为一种特别有价值的工具，因为它是 应用于三维ECM中的细胞，这是大多数细胞的自然状态。当前的3D TFM 使用光学图像生成3D几何模型的关键步骤挑战了方法 细胞周围的基质，并根据微珠的位移估计来推断细胞术 嵌入矩阵中。在这些步骤中发生的近似导致计算障碍的重大错误 这反过来导致了错误的生物学结论。因此，迫切需要更准确和 高分辨率3D TFM技术。 拟议研究的长期目标是改进和自动化3D TFM流程，以便它 可以有效地用于回答机械生物学问题并设计新的治疗干预措施。这 将通过（a）将高级细分和网格生成技术应用于光学 图像生成一个细胞周围基质的3D几何模型和有限元网格，以及（b） 通过开发和实施新算法以确定细胞障碍的空间分布 测量的微珠位移，同时考虑了基质的非线性弹性响应。这些 发展将通过基准研究验证，其实用性将通过量化来证明 嵌入合成细胞外基质中的癌细胞施加的牵引力。