A Biophotonics Platform for Mechanotransduction and Metabolic Microscopy

用于机械转导和代谢显微镜的生物光子学平台(M3) 项目总结：(30行) 机械转导已成为生物功能的主要调节因子之一。而当 细胞机械转导的下游效应已经被广泛研究，相互作用 细胞机械转导与新陈代谢之间的关系尚不清楚。这 该提案代表了机械生物学专家之间的多学科合作， 激光与组织的相互作用，以及先进的荧光显微镜方法开发了一种独特的 生物光子学技术平台，使脉冲力能够精确应用于2-D 用于动态测量细胞的细胞培养、3D组织培养和活体动物模型 机械敏感性和细胞机械信号，并将它们与代谢状态相关联， 膜流动性、信使RNA(MRNA)表达以及细胞/基质组成， 形态、组织和僵硬。这将通过一种独特的集成实现 激光扫描共聚焦用于细胞/组织刺激的脉冲激光微束照射 (LSCM)、多光子(MPM)、荧光寿命成像(FLIM)显微镜和多光子 光谱成像模式。我们将在一年内验证和演示此平台的功能 各种复杂程度和材料不同的二维细胞培养和三维组织培养系统 构图和硬度。拟议的技术开发将使独特的 测量和检查对机械敏感性的动态细胞相互作用的能力 信号、机械信号和其他生物系统特征，包括细胞 代谢、信使核糖核酸表达和细胞外基质重塑。建议的技术 平台在正常组织发育和疾病的研究中具有广泛的应用前景 高通量药物筛选。

A Biophotonics Platform for Mechanotransduction and Metabolic Microscopy (M3) Project Summary: (30 lines) Mechanotransduction has emerged as one of the main regulators of biological function. While the downstream effects of cellular mechanotransduction have been widely studied, the interplay between cellular mechanotransduction and metabolism remains relatively unexplored. This proposal represents a multi-disciplinary collaboration between specialists in mechanobiology, laser-tissue interactions, and advanced fluorescence microscopy methods to develop a unique biophotonics technology platform that enables the precise application of impulsive force to 2-D cell cultures, 3-D tissue cultures, and live animal models to dynamically measure cellular mechanosensitivity and cellular mechanosignaling and correlate these with metabolic state, membrane fluidity, messenger RNA (mRNA) expression, as well as cell/matrix composition, morphology, organization, and stiffness. This will be achieved through a unique integration pulsed laser microbeam irradiation used for cell/tissue stimulation with laser scanning confocal (LSCM), multi-photon (MPM), fluorescence lifetime imaging (FLIM) microscopy, and multi- spectral imaging modalities. We will verify and demonstrate the capabilities of this platform in a variety of 2-D cell culture and 3-D tissue culture systems of varying complexity and material composition and stiffness. The proposed technology development will enable a unique capability to measure and examine the dynamic cellular interplay of sensitivity to mechanical cues, mechanosignalling, and other biological system characteristics including cellular metabolism, mRNA expression, and extracellular matrix remodeling. The proposed technology platform has broad applications for the study of normal tissue development and disease and for high-throughput drug screening.