High-End Biological Atomic Force Microscope

项目概要 这是2020年“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， 关于力学对生物系统的影响的研究领域不断发展。学习机械生物学 在分子、细胞和组织水平上，研究人员需要一种能够测量和控制分子、细胞和组织力学的工具 样品，同时监测样品内的生物活性。本提案中要求的文书是 将原子力显微镜 (AFM) 放置在荧光显微镜顶部的集成系统， 提供独特的功能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、高分辨率机械属性映射、视频 速率成像和纳米流体集成探针系统。荧光显微镜包括一个旋转 圆盘共焦、全内反射荧光 (TIRF) 和微分干涉对比 (DIC)。 PI 各制造商将整合这两个系统，建立一个组合平台（Bio-AFM）， 可以实时执行先进的同步机械和光学测量，提供新颖的 多种目标生物系统的机械生物学研究方法。 访问 Bio-AFM 将立即推进 5 个用户中的 16 个用户（10 个主要用户和 6 个次要用户）的研究 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、海军和陆军研究项目资助。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的生物物理学（3个项目）、 生物医用材料（3个项目）、力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）和诊断生物传感（1 个项目）。 Bio-AFM 将安置在一个完善的核心设施中，每个有需要的人都可以使用它 它。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很容易适应 容纳生物原子力显微镜。 PI 是表面科学核心设施的创始主任（拥有 20 多名 多年的 AFM 研究经验）并将负责监督该设施的所有 Bio-AFM 活动。 与表面科学核心设施中的所有其他仪器一样，Bio-AFM 将得到 财务、安全、外展、IT、拨款管理和行政方面的机构。鉴于 专业知识、基础设施和机构支持，所要求的 Bio-AFM 不仅将使最初的 14 个用户受益， 同时也为无数未来的用户和生物医学研究延伸的新项目提供了新的机会 远远超出了资助期限的结束时间。

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.