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

描述（由申请人提供）：该项目的特点是集成了先进的光子技术和微流体技术，以应对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生物化学和生物物理刺激的复杂混合物来严格调节细胞功能。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。需要改进的一个领域是细胞内弹性的非侵入性测量。弹性介导细胞内部的力的传递和细胞区域在施加的力下所经历的变形。然而，目前用于细胞/ECM弹性测量的技术限于点样品分析或需要接触。这些都是重要的限制，因为细胞是异质的，改变其性质后，机械扰动，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。布里渊细胞显微术 可以在3D培养物中以高分辨率、非扰动、无接触地绘制细胞内弹性。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和细胞内机械转导的检测共存。与微流体平台的集成将使微环境条件的严格控制成为可能。在仪器验证之后，我将专注于乳腺癌细胞迁移。基于初步的数据和文献证据，我制定并将测试的假设，即细胞内弹性介导migratio，即最佳的细胞模量和弹性极化的迁移机械的机械要求，并可以用来解释增强的运动性转移细胞相比，他们的非癌对应。除了细胞迁移研究之外，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定量指标，将细胞-微环境机械相互作用与细胞行为联系起来。这个K25奖将使候选人的过渡到细胞生物力学和机械生物学的研究，通过补充他在光学技术开发方面的专业知识，通过正式的课程，与导师的互动和动手实验室培训，在细胞生物学，微流体和道德研究行为方面进行必要的培训。

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.