Combined Atomic Force and Total Internal Reflection Fluorescence Microscopy for Biological Research

用于生物学研究的原子力和全内反射荧光显微镜相结合

• 批准号: 9604785
• 负责人: Ashutosh Chilkoti
• 金额: $ 13.22万
• 依托单位: Duke University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-15 至 1999-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9604785&HistoricalAwards=false
• 关键词: Combined; Atomic; Force; Total; Internal

The Center for Cellular and Biosurface Engineering at Duke University requests funding to purchase atomic force microscopy (AFM) instrumentation for the shared use of faculty within the Center. No such instrumentation currently exists within the Center for shared use. A significant number of the core investigators have used atomic force microscopes at other institutions. The requested Digital Instruments MultiMode/BioscopeTM AFM allows simultaneous optical microscopy and AFM imaging. We propose to further extend the capabilities of the BioscopeTM AFM by incorporating total internal reflection fluorescence microscopy (TIRFM) into its optics. This instrument will enhance the research of six members in the interdisciplinary Center for Cellular and Biosurface Engineering (from four departments) in the following research projects: (1) characterization of crystalline, two-dimensional bacterial s-layer protein arrays and nanopatterned protein arrays directed by the s-layer nanotemplates; (2) elucidation of the mechanism of mechanical signal transduction in musculoskeletal cells; (3) characterization of photopatterned antibody arrays designed for a multianalyte integrated optical waveguide sensor; (4) quantitative imaging of endothelial cell attachment and spreading processes on novel, micropatterned cell-adhesive ligand substrates; (5) characterization of a DNA chip: multiplexed, spatially-localized oligonucleotide arrays on silicon substrates; (6) analysis of motor proteins by AFM; and (7) development of a biophysical model to relate protein-ligand recognition energetics to AFM-measured intermolecular forces. The acquisition of an AFM will also significantly enhance the educational mission of the Center for Cellular and Biosurface Engineering at Duke University. The Center was the recent recipient of a special opportunity award from the Whitaker Foundation for educational enhancements to the program at both the undergraduate and graduate levels. The Center is committed to developing innovative laboratory classes that utilize contemporary experimental techniques to understand, quantify, and visualize the fundamental forces, stresses, and recognition processes at the molecular and cellular levels that are central to biological phenomena. The ability of the atomic force microscope to investigate these phenomena in a biologically-relevant milieu in real time combined with its relative ease of use makes it an ideal experimental technique for a number of the laboratory components of a new graduate course under development.

杜克大学细胞和生物表面工程中心申请资金购买原子力显微镜(AFM)仪器，供中心内的教职员工共享使用。目前，共享使用中心内没有这样的仪器。相当数量的核心调查人员在其他机构使用了原子力显微镜。所要求的数字仪器多模/Bioscope eTM AFM允许同时进行光学显微镜和AFM成像。我们建议通过将全内反射荧光显微镜(TIRFM)引入其光学系统来进一步扩展Bioscope TM AFM的能力。该仪器将加强细胞和生物表面工程跨学科中心(来自四个部门)的六名成员在下列研究项目中的研究：(1)S纳米模板指导的结晶二维细菌S层蛋白质阵列和纳米蛋白质阵列的表征；(2)阐明肌肉骨骼细胞中机械信号转导的机制；(3)用于多分析集成光波导传感器的光修饰抗体阵列的表征；(4)内皮细胞在新型微图案化细胞-粘附性配体和基质上附着和扩散过程的定量成像；(5)DNA芯片的表征：硅衬底上多路、空间定位的寡核苷酸阵列；(6)用原子力显微镜分析马达蛋白质；(7)建立生物物理模型，将蛋白质-配体识别能量学与原子力显微镜测量的分子间力联系起来。收购AFM还将显著增强杜克大学细胞和生物表面工程中心的教育使命。该中心最近获得了惠特克基金会颁发的特别机会奖，以表彰该项目在本科生和研究生层面的教育改进。该中心致力于开发创新的实验室课程，利用现代实验技术来理解、量化和可视化分子和细胞水平上的基本力量、压力和识别过程，这些过程是生物现象的核心。原子力显微镜在与生物相关的环境中实时研究这些现象的能力，加上其相对易用性，使其成为正在开发的新研究生课程的许多实验室组件的理想实验技术。