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

项目描述（申请人提供）：本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解决的重大挑战。细胞微环境通过提供复杂的生物化学和生物物理刺激来调节细胞功能。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，在开发用于机械刺激和力响应的单细胞工具方面取得了巨大进展。一个需要改进的领域是细胞内弹性的非侵入性测量。弹性调节细胞内力的传递和细胞区域在外力作用下的变形。然而，目前的电池/ECM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，并且需要在3D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满足的需求。布里渊细胞显微镜

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.