MRI: Track 1 Acquisition of a System for Integrated Confocal Microscopy and Mechanical Interrogation

这一重大研究仪器(MRI)奖支持收购三种商用仪器，这三种仪器将合并在一起创造一种新的仪器。将获得的三种仪器是共焦显微镜、原子力显微镜(AFM)和激光切割机。这种共聚焦显微镜将能够对活细胞和组织以及软凝胶和其他软材料进行快速、高分辨率的光学成像。原子力显微镜将能够在这些样品上定义良好的位置施加良好控制的力，以测量它们的机械性能。此外，原子力显微镜还可以用来对样品表面进行比共焦显微镜更高分辨率的成像。激光切割器将用于在生物组织或纤维网络处于拉伸状态时切割它们，因此可以通过测量切割后的反冲来测量结构中的张力。通过共同努力，这些仪器的组合将能够确定生物细胞和组织中形成的机械特性，以及生物细胞和组织如何对来自其环境的一系列机械输入做出反应。虽然众所周知，生物学和力学之间的相互作用在广泛的基本生物过程中是重要的，但用于探索不同类型相互作用的特定工具一直受到限制。因此，该仪器的获得将极大地促进对生物系统和在组成、结构和/或功能上与生物系统相似的非生物系统的基本科学理解。该仪器的获得还将推动材料工程的发展，这些材料与生物系统对接，以期望的方式调整生物反应。这将通过生物医学的多个领域的进步造福社会。该仪器将为处于教育和职业生涯各个阶段的科学家和工程师提供新的培训和研究机会。具体的努力将集中在使职业生涯早期的研究人员和那些代表不足的群体成员受益。一个高分辨率、高速共聚焦显微镜、一个专为处理“湿”生物和软物质样本而设计的原子力显微镜，以及一个激光切割器将被组合在一起，创造出一种称为“机械显微镜”的仪器。该仪器将用于研究机械生物学，包括表征细胞细胞骨架和细胞外基质的纤维网络，单细胞真核和原核生物机械生物学(特别强调信号和分化)，以及组织和生物水平的机械生物学(特别强调结缔组织和形态发生)。该仪器还将用于研究生物-非生物界面的相互作用，重点是光响应型动态水凝胶。此外，该仪器将用于开发新的软材料，包括活性软凝胶、肽原纤维和刺激响应性软材料，这些软材料将基因工程菌作为调节凝胶特性的功能组件。共焦显微镜将用于成像和福斯特/荧光共振能量转移(FRET)测量。原子力显微镜将用于控制力的施加以及杨氏模数和存储和损耗模数的测量。激光切割器将用于烧蚀纤维网络、细胞骨架和组织，并用作交联动态水凝胶的光刺激。该项目由重大仪器研究计划(MRI)和土木、机械和制造创新(CMMI)部门的生物力学和机械生物学计划(BMMB)联合资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This Major Research Instrumentation (MRI) award supports the acquisition of three commercially-available instruments that will be combined to create a new instrument. The three instruments to be acquired are a confocal microscope, an atomic force microscope (AFM), and a laser cutter. The confocal microscope will be able to do fast, high-resolution optical imaging of living cells and tissues as well as soft gels and other soft materials. The AFM will be able to apply well-controlled forces at well-defined locations on these samples, to measure their mechanical properties. Additionally, the AFM can also be used to do higher-resolution imaging of sample surfaces than the confocal light microscope. The laser cutter will be used to cut biological tissues or fiber networks while they are under tension, so that the amount of tension in the structure can be measured by measuring the recoil following the cut. Working together, the combination of these instruments will allow determination of the mechanical properties that develop in biological cells and tissues and how biological cells and tissues respond to a range of mechanical inputs from their environments. Although it is well-known that the interplay between biology and mechanics is important in a wide range of essential biological processes, specific tools for probing the different types of interplay have been limited. Therefore, the acquisition of this instrument will substantially advance basic scientific understanding of biological systems and of non-biological systems that resemble biological systems in composition, structure, and/or function. Acquisition of this instrument will also advance the engineering of materials that interface with biological systems to tune biological response in the desired way. This will benefit society through advances in multiple areas of biomedicine. The instrument will offer new training and research opportunities to scientists and engineers at all stages of their education and career. Specific efforts will be focused on benefiting early-career researchers and those who are members of under-represented groups.A high-resolution, high-speed confocal microscope, an AFM designed for work with “wet” biological and soft-matter samples, and a laser cutter will be combined to create one instrument, termed the “mechanoscope.” This instrument will be used to investigate mechanobiology, including the fiber networks that characterize the cellular cytoskeleton and the extracellular matrix, single-cell eukaryotic and prokaryotic mechanobiology (with a particular emphasis on signaling and differentiation), and tissue- and organism-level mechanobiology (with a particular emphasis on connective tissue and morphogenesis). This instrument will also be used to investigate interactions at the biotic-abiotic interface, focusing on photo-responsive dynamic hydrogels. Further, the instrument will be used in the development of new soft materials, including active soft gels, peptide fibrils, and stimuli-responsive soft materials that incorporate genetically engineered bacteria as a functional component for tuning gel properties. Confocal microscopy will be used for imaging and Forster/fluorescence resonance energy transfer (FRET) measurements. AFM will be used for controlled force application and for measurement of Young’s moduli and storage and loss moduli. The laser cutter will be used for ablation of fiber networks, cellular cytoskeletons, and tissues, and as a photo-stimulus for crosslinking dynamic hydrogels.This project is jointly funded by the Major Instrumentation Research Program (MRI) and the Biomechanics and Mechanobiology Program (BMMB) in the division of Civil, Mechanical and Manufacturing Innovation (CMMI).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.