Imaging cellular biomechanics on-chip in 2D and 3D microenvironments

2D 和 3D 微环境中的片上细胞生物力学成像

• 批准号: 9057538
• 负责人: Giuliano Scarcelli
• 金额: $ 14.23万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9057538
• 关键词: Acoustics; Actins; Anisotropy; Area; Award; Behavior; Benchmarking; Biochemical; Biological; Biomechanics; Breast Cancer Cell; Calibration; Cell Line; Cell physiology; Cells; Cellular Structures; Cellular biology; Collagen; Complement; Complex Mixtures; Data; Detection; Development Plans; Elasticity; Environment; Ethics; Exhibits; Extracellular Matrix; Focal Adhesions; Frequencies; Gene Expression; Goals; Health; Hour; Hydrogels; Image; Laboratories; Light; Literature; MCF10A cells; MCF7 cell; MDA MB 231; Mammary gland; Maps; Measurement; Measures; Mechanical Stimulation; Mechanics; Mediating; Mentors; Mentorship; Metastatic breast cancer; Methods; Microfluidics; Microscopy; Modeling; Modification; Morphogenesis; Morphologic artifacts; Nature; Neoplasm Metastasis; Optics; Pathway interactions; Physics; Property; Reading; Regulation; Research; Resolution; Rheology; Role; Sampling; Science; Signal Transduction; Speed; Stimulus; System; Techniques; Technology; Testing; Time; Tissues; Training; Validation; Variant; angiogenesis; base; biological research; career development; cell behavior; cell motility; cellular imaging; experience; field study; imaging modality; improved; in vivo; instrument; laboratory experience; light scattering; medical schools; migration; novel; optical imaging; photonics; physical science; polyacrylamide gels; programs; reconstitution; response; technology development; three dimensional cell culture; tool; transmission process

DESCRIPTION (provided by applicant): This project features the integration of advanced photonic technology and microfluidics to attack a major unmet challenge in cell biomechanics. The cellular microenvironment critically regulates cellular function by providing a complex mixture of biochemical and biophysical stimuli. Among the components of the cell-microenvironment interaction, the role of biomechanical factors is recognized to be crucial. In recent years, tremendous progress has been achieved in developing single-cell tools for mechanical stimulation and force response. One area of needed improvement is the non-invasive measurement of intracellular elasticity. Elasticity mediates the transmission of forces inside the cell and the deformation experienced by cell regions under an applied force. However, current technology for cell/ECM elasticity measurements is limited to point-sample analysis or requires contact. These are important limitations since cells are heterogeneous, alter their properties upon mechanical perturbation, and need to be studied in 3D microenvironments. This project will develop an all-optical approach to this unmet need. Brillouin cellular microscopy can map the intracellular elasticity at high resolution, non-perturbatively, without contact in 3D cultures. Brillouin information on cell elasticity will be co-located with fluorescent-based detection of cytoskeletal components and intracellular mechanotransduction. Integration with microfluidic platforms will enable tight control of microenvironment conditions. After instrument validation, I will focus on breast cancer cell migration. Based on preliminary data and literature evidence, I formulated and will test the hypothesis that intracellular elasticity mediates migratio, namely that optimal cell modulus and elasticity polarization are mechanical requirements of the migration machinery and can be used to explain the enhanced motility exhibited by metastatic cells compared to their non-cancerous counterparts. Beyond the cell migration studies, the novel instrumental platform developed and validated during this award, will be broadly applicable as it provides unique quantitative metrics to relate cell- microenvironment mechanical interaction to cell behavior. This K25 award would enable the candidate's transition to research in cellular biomechanics and mechanobiology by complementing his demonstrated expertise in optical technology development with the necessary training in cell biology, microfluidics and ethical research conduct through formal coursework, interaction with mentors and hands-on laboratory training.

描述(由申请者提供)：该项目将先进的光子技术和微流体技术相结合，以解决细胞生物力学中尚未满足的主要挑战。细胞微环境通过提供生化和生物物理刺激的复杂混合物来关键地调节细胞功能。在细胞-微环境相互作用的组成部分中，生物力学因素的作用被认为是至关重要的。近年来，用于机械刺激和力响应的单细胞工具的开发取得了巨大的进步。需要改进的一个领域是细胞内弹性的非侵入性测量。弹性调节细胞内部的力的传递，以及细胞区域在外力作用下所经历的变形。然而，目前的细胞/细胞外基质弹性测量技术仅限于点样分析或需要接触。这些都是重要的限制，因为细胞是异质的，在机械扰动下会改变它们的性质，需要在三维微环境中进行研究。该项目将开发一种全光学方法来满足这一未得到满足的需求。布里渊细胞显微镜 可以在3D培养中以高分辨率、非微扰、无接触的方式绘制细胞内弹性图。关于细胞弹性的布里渊信息将与基于荧光的细胞骨架成分检测和细胞内机械转导共同定位。与微流控平台的集成将实现对微环境条件的严格控制。在仪器验证后，我将重点研究乳腺癌细胞的迁移。基于初步数据和文献证据，我提出并将检验细胞内弹性介导迁移的假设，即最佳细胞弹性系数和弹性极化是迁移机制的力学要求，可以用来解释转移细胞与非癌细胞相比表现出的增强的运动性。除了细胞迁移研究，在该奖项期间开发和验证的新型仪器平台将广泛适用，因为它提供了独特的量化指标，将细胞-微环境机械相互作用与细胞行为联系起来。这项K25奖项将使候选人能够过渡到细胞生物力学和机械生物学的研究，通过正规的课程作业、与导师的互动和动手实验室培训，补充他在光学技术开发方面所展示的专业知识，以及在细胞生物学、微流体学和伦理研究方面的必要培训。