High-End Biological Atomic Force Microscope

项目摘要 这是对2020年度《高端荧光生物原子力显微镜》提案的重新提交 NIH S10计划(申请号：1 S10 OD030401-01)专注于机械生物学研究，快速 生物系统中力学影响的研究领域日益扩大。在大学攻读机械生物学 分子、细胞和组织水平，研究人员需要一种工具来测量和控制 在监测样本内的生物活动的同时，对样本进行监测。本提案所要求的文书是 一种集成系统，将原子力显微镜(AFM)放置在荧光显微镜的顶部， 提供独特的能力来精确测量和施加力并同时可视化生物 现象。AFM平台提供力测量、高分辨率机械特性映射、视频- 速率成像，以及纳米流体集成探头系统。该荧光显微镜包括一个旋转的 圆盘共聚焦、全内反射荧光(TIRF)和差分干涉对比(DIC)。《少年派》 而各自的制造商将整合这两个系统，以建立一个组合平台(Bio-AFM)， 可实时执行先进的同步机械和光学测量，提供新颖的 对多种目标生物系统进行机械生物学研究的方法。 使用Bio-AFM将立即推进对16个用户(10个主要用户和6个次要用户)的研究，从5个 大学和11个系，包括由NIH资助的9个用户(过去5年为1200万美元)和6个用户 由美国国家科学基金会、空军、海军和陆军研究项目资助。这些初步项目的进展将会取得进展 神经元疾病研究(6个项目)，动脉粥样硬化和微血管疾病的生物物理学研究(3个项目)， 生物医学材料(3个项目)、机械传感及其在免疫学中的应用(2个项目)、骨相关 疾病(1个项目)和生物传感诊断(1个项目)。 Bio-AFM将被安置在一个完善的核心设施中，每个需要的人都可以接触到它 它。表面科学核心设施目前为来自70多个研究项目的230多个终端用户提供服务 组。现有的仪器培训、操作和管理基础设施很容易适应 安装Bio-AFM。PI是表面科学核心设施的创始主任(拥有20多名 多年的AFM研究经验)，并将负责监督该设施的所有生物AFM活动。 与表面科学核心设施中的所有其他仪器一样，生物原子力显微镜将得到 在财务、安全、外展、信息技术、赠款管理和行政方面的机构。给定 除了专业知识、基础设施和机构支持外，所要求的生物原子力显微镜不仅将使最初的14名用户受益， 而且还为无数未来的用户提供了新的机会，为生物医学研究扩展提供了新的项目 远远超过授权期结束的时间。

Project Summary This is a re-submission of the proposal “High-End Fluorescence Biological Atomic Force Microscope” of 2020 NIH S10 program (Application Number: 1 S10 OD030401-01) focusing on mechanobiological studies, a rapidly growing field of research on the influence of mechanics in biological systems. To study mechanobiology at the molecular, cellular, and tissue levels, researchers need a tool that can measure and control the mechanics of samples while monitoring biological activities within the samples. The instrument requested in this proposal is an integrated system that positions an atomic force microscope (AFM) on top of a fluorescence microscope, providing unique capabilities to precisely measure and apply forces and simultaneously visualize biological phenomena. The AFM platform offers force measurements, high-resolution mechanical property mapping, video- rate imaging, and a nano-fluidics integrated probe system. The fluorescence microscope includes a spinning disk confocal, total internal -reflection fluorescence (TIRF), and differential interference contrast (DIC). The PI and the respective manufacturers will integrate the two systems to set up a combined platform (Bio-AFM) that can perform advanced and simultaneous mechanical and optical measurements in real time, providing novel approaches for mechanobiological studies of a broad variety of target bio-systems. Access to the Bio-AFM will immediately advance the research of 16 users (10 major and 6 minor) from 5 universities and 11 departments, including 9 users funded by the NIH (> $12M in the past 5 years) and 6 users funded by NSF, Air Force, Navy, and Army research programs. Progress in those initial projects will advance research on neuron disorders (6 projects), the biophysics of atheroma and microvascular disease (3 projects), biomedical materials (3 projects), mechano-sensing and its application in immunology (2 projects), bone-related disorders (1 project), and bio-sensing for diagnostics (1 project). The Bio-AFM will be housed in a well-established core facility where it will be accessible to everyone that needs it. The Surface Science Core Facility currently serves more than 230 end users from more than 70 research groups. The existing infrastructure for instrument training, operation, and management is easily adaptable to accommodate the Bio-AFM. The PI is the Surface Science Core Facility’s founding Director (with more than 20 years of AFM research experience) and will be responsible for overseeing all Bio-AFM activities at the facility. The Bio-AFM, like all other instruments in the Surface Science Core Facility, will be fully supported by the institution with regard to finances, safety, outreach, IT, grant management, and administration. Given the expertise, infrastructure, and institutional supports, the requested Bio-AFM will not only benefit the initial 14 users, but also provide new opportunities to countless future users and new projects for biomedical research extending far beyond the end of the grant period.