Bio-AFM for combined light and atomic force imaging

用于组合光和原子力成像的生物原子力显微镜

• 批准号: 9074870
• 负责人: Alexander R Dunn
• 金额: $ 51.33万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2017-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9074870
• 关键词: Acoustics; Area; Biocompatible Materials; Biological; Biophysical Process; Carbon Dioxide; Cardiac Myocytes; Cell division; Cells; Communities; Cues; Discipline; Disease; Engineering; Esthesia; Funding; Head; Health; Image; Light; Maps; Measurement; Mechanics; Mediating; Methodology; Microbe; Microscope; Microscopy; Modeling; Molecular; Optics; Organoids; Performance; Phase; Proteins; Publishing; Qualifying; Regenerative Medicine; Research; Research Personnel; Research Project Grants; Resolution; Sampling; Signal Pathway; Signal Transduction; Staging; Stem cells; Stimulus; Students; Subcellular structure; Surface; T-Lymphocyte; Techniques; United States National Institutes of Health; Universities; active control; base; design; environmental change; familial dilated cardiomyopathy; innovation; instrument; macromolecule; novel; optical imaging; research study; vibration

 DESCRIPTION (provided by applicant): We propose to acquire a combined atomic force and optical microscope that will be equally useful for dynamic biological sample and soft/wet material research (Bio-AFM). The BioScope Resolve-Bio AFM (Bruker, Inc.) head and AxioObserver Z1 epifluorescence microscope (Zeiss, Inc.) base we are requesting has a versatile, modular design that combines the sub-molecular resolution imaging and precision force measurements of AFM with a broad range of optical microscopy techniques for simultaneous, correlative AFM and optical microscopy. This is a high performance Bio-AFM consisting of the following components: 1) Bio-AFM head equipped with an extended z-range head, Petri dish holder and stage heater, CO2 control, active vibration control and acoustic enclosure. 2) Inverted epifluorescence optical microscope on which to mount the AFM head; provides automated, diffraction limited resolution, wide-field epifluorescence and brightfield /phase imaging capabilities with piezo z-drive, sCMOS camera, and splitting optics, needed for correlative biological imaging studies utilizing AFM. The Bio- AFM and its requested accessories will serve first the specific needs of the NIH funded research projects described in this proposal and then the anticipated needs of the greater Stanford University and surrounding research community. This instrument will enable innovative experiments that will allow high resolution force measurements and mapping over the surface of soft materials, cells and other biological material. These force measurements will be correlated with macromolecules, proteins and subcellular structures as cells sense and respond to mechanical cues and environmental changes via epifluorescence, brightfield and phase contrast optical imaging. These otherwise not possible combinations of imaging experiments will result in dramatic and rapid progress in important health-related research topics in a variety of disciplines and interdisciplinary areas including the molecular and biophysical mechanisms underlying sensation of mechanical stimuli, engineering organoid cultures for regenerative medicine research, analysis of stem cell derived cardiomyocyte contractility and modeling familial dilated cardiomyopathy, correlating proteins involved in cell division with force distribution in dividing microbes and the manipulatio of T cell signaling pathways to control immunologically-mediated diseases. Stanford researchers, students, and staff are well-qualified - and eager - to carry out the research described here, to use the Bio-AFM to further these and other research efforts, to develop and publish novel methodologies for Bio-AFM, and to maintain the instrument within a shared imaging facility that will enable additional research.

 描述（由申请人提供）：我们建议获得一个组合的原子力和光学显微镜，这将是同样有用的动态生物样品和软/湿材料的研究（生物原子力显微镜）。BioScope Resolve-Bio AFM（Bruker，Inc.）头和AxioObserver Z1落射荧光显微镜（Zeiss，Inc.）我们所要求的一个多功能的模块化设计，将AFM的亚分子分辨率成像和精确力测量与广泛的光学显微镜技术相结合，用于同时进行相关的AFM和光学显微镜。这是一个高性能的生物原子力显微镜，由以下组件组成：1）生物原子力显微镜头配备了一个扩展的z范围头，培养皿保持器和阶段加热器，二氧化碳控制，主动振动控制和隔音罩。2)安装AFM头的倒置落射荧光光学显微镜;提供自动化、衍射极限分辨率、宽场落射荧光和亮场/相位成像能力，具有压电z驱动、sCMOS相机和分光光学器件，用于利用AFM进行相关生物成像研究。生物原子力显微镜及其要求的配件将首先服务于本提案中描述的NIH资助的研究项目的具体需求，然后服务于更大的斯坦福大学和周边研究社区的预期需求。该仪器将使创新的实验，将允许高分辨率的力测量和映射在软材料，细胞和其他生物材料的表面。这些力的测量将与大分子，蛋白质和亚细胞结构相关，因为细胞通过落射荧光，明场和相衬光学成像来感知和响应机械线索和环境变化。这些成像实验的其他不可能的组合将导致各种学科和跨学科领域中重要的健康相关研究主题的戏剧性和快速进展，包括机械刺激感觉的分子和生物物理机制，用于再生医学研究的工程类器官培养物，干细胞衍生的心肌细胞收缩性的分析和建模家族性扩张型心肌病，将参与细胞分裂的蛋白质与分裂微生物中的力分布相关联，以及操纵T细胞信号传导途径以控制免疫介导的疾病。斯坦福大学的研究人员、学生和工作人员都有资格--并且渴望--进行这里描述的研究，使用生物原子力显微镜来推进这些和其他研究工作，开发和发布生物原子力显微镜的新方法，并在一个共享的成像设施内维护仪器，这将使更多的研究成为可能。