An Enhanced Combinatorial Super-Resolution Microscopy Platform

增强型组合超分辨率显微镜平台

• 批准号: RTI-2016-00511
• 负责人: Yip, Christopher
• 金额: $ 10.93万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=591473
• 关键词: Enhanced; Combinatorial; Super; Resolution; Microscopy

RTI funding will be used to enhance the spatial, temporal, and spectral resolution of an exceptionally capable super-resolution combinatorial microscopy platform that combines the sub-nanometer 3-D resolution of scanning probe microscopy with super-resolution fluorescence microscopy. This latter technique (and its variants) was recently awarded the 2014 Nobel Prize. The platform to be upgraded is unique in Canada with capabilities lacking in commercial systems. Comprising a 15 year old scanning probe microscope (SPM), a home-built, self-funded 4-laser dual-camera total internal reflection microscope, and a 4-laser confocal microscope, this platform is critical to multiple successful inter-disciplinary, -faculty, and -university collaborations, broadly supporting projects in structural biology, biophysics, and engineering. The present proposal seeks funds to replace the current SPM with a state-of-the-art system that has enhanced scan performance, significantly reduced noise, and higher sensitivity and resolution. This replacement is critically required as the current SPM is no longer supported by the vendor, and has become increasingly unreliable. Components cannot be sourced as they are no longer being manufactured leaving eBay and other auction sites as the only alternatives for parts. We are also seeking funding for a 561 nm laser to be integrated into the existing optical train (405 nm / 473 nm / 532 nm / 637 nm laser lines). This line will excite many of the key fluorophores for super-resolution imaging. The open-beam design of our optical train allows for ready control over phase, alignment, polarization, and beam shape, attributes that are not as easily manipulated within commercial systems.

RTI的资金将用于提高超分辨率组合显微镜平台的空间、时间和光谱分辨率，该平台结合了扫描探针显微镜的亚纳米三维分辨率和超分辨率荧光显微镜。后一种技术（及其变体）最近获得了2014年诺贝尔奖。升级的平台在加拿大是独一无二的，具有商业系统所缺乏的能力。该平台包括一台已有15年历史的扫描探针显微镜（SPM）、一台自建、自筹资金的4激光双摄像头全内反射显微镜和一台4激光共聚焦显微镜，对多个成功的跨学科、院系和大学合作至关重要，广泛支持结构生物学、生物物理学和工程学领域的项目。目前的提案寻求资金，用最先进的系统取代目前的SPM，该系统具有增强的扫描性能，显着降低噪音，更高的灵敏度和分辨率。这种替换是非常必要的，因为当前的SPM不再得到供应商的支持，并且变得越来越不可靠。零件无法采购，因为它们不再生产，eBay和其他拍卖网站成为零件的唯一替代品。我们还在寻求资金，将561纳米激光器集成到现有的光学系列（405纳米/ 473纳米/ 532纳米/ 637纳米激光线）中。这条线将激发许多关键的荧光团超分辨率成像。我们的光学列车的开光束设计允许对相位、对准、偏振和光束形状进行随时控制，这些属性在商业系统中不容易操作。