High-End Biological Atomic Force Microscope

项目摘要 这是2020年“高端荧光生物原子力显微镜”提案的再次提交 NIH S10计划（申请号：1 S10 OD 030401 -01）专注于机械生物学研究， 不断增长的研究领域的影响力学在生物系统。去研究机械生物学 在分子、细胞和组织水平上，研究人员需要一种工具来测量和控制细胞的力学。 同时监测样品中的生物活性。本提案所要求的工具是 将原子力显微镜（AFM）定位在荧光显微镜顶部的集成系统， 提供独特的能力，精确测量和施加力，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- 速率成像和纳米流体集成探针系统。荧光显微镜包括旋转 圆盘共聚焦、全内反射荧光（TIRF）和微分干涉对比（DIC）。的PI 而各自的制造商将整合两个系统，建立一个组合平台（Bio-AFM）， 可以在真实的时间内进行先进的同时机械和光学测量， 为各种各样的目标生物系统的机械生物学研究的方法。 访问生物原子力显微镜将立即推进研究的16个用户（10个主要和6个次要）从5 大学和11个部门，包括9个由NIH资助的用户（过去5年超过1200万美元）和6个用户 由美国国家科学基金会、空军、海军和陆军研究计划资助。这些初步项目的进展将取得进展 神经元疾病研究（6个项目），动脉粥样硬化和微血管疾病生物物理学（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）和用于诊断的生物传感（1个项目）。 生物原子力显微镜将被安置在一个完善的核心设施，在那里它将是每个人都需要访问 了表面科学核心设施目前为来自70多个研究领域的230多个最终用户提供服务。 组现有的仪器培训、操作和管理基础设施可轻松适应 适应生物原子力显微镜PI是表面科学核心设施的创始主任（拥有20多个 年的AFM研究经验），并将负责监督所有生物原子力显微镜活动的设施。 生物原子力显微镜，像所有其他仪器在表面科学核心设施，将完全支持 在财务、安全、外联、信息技术、赠款管理和行政管理方面，鉴于 专业知识，基础设施和机构支持，所要求的生物原子力显微镜将不仅有利于最初的14个用户， 也为无数未来的用户提供了新的机会，为生物医学研究提供了新的项目， 远远超过了补助金期限的结束。

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.