A FRAP on TIRF superresolution microscopy system for bioimaging

用于生物成像的 TIRF 超分辨率显微镜系统上的 FRAP

• 批准号: RTI-2016-00097
• 负责人: Wasteneys, Geoffrey
• 金额: $ 7.06万
• 依托单位: University of British Columbia
• 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=591342
• 关键词: FRAP; TIRF; superresolution; microscopy; system

The tremendous power of fluorescence microscopy was recently recognized with the 2014 Nobel Prize in Chemistry to three scientists who have masterminded super resolution imaging. Although hailed as a quantum leap in live cell imaging, there are few labs worldwide who can actually use super resolution in living cells. Other modes of fluorescence imaging are now available that can improve resolution and also obtain quantitative data on the dynamics of cellular processes in real time. This project proposes to combine two such imaging modes, total internal reflectance fluorescence (TIRF) and fluorescence redistribution after photobleaching (FRAP) on one microscope. Specifically, an existing multi-laser TIRF microscope will be retrofitted with the necessary hardware to enable FRAP to be conducted and to generate data on the dynamic turnover of proteins at the cell surface. This new tool will be indispensable for quantifying the turnover of enzyme complexes that synthesize the worlds most abundant biopolymer cellulose, as well as membrane receptors, transporters and other proteins complexes situated at the plasma membrane.

荧光显微镜的巨大力量最近被2014年诺贝尔化学奖授予了三位策划超分辨率成像的科学家。虽然被誉为活细胞成像的一次飞跃，但世界上很少有实验室能够真正在活细胞中使用超分辨率。其他模式的荧光成像，现在可以提高分辨率，并获得定量数据的动态细胞过程中的真实的时间。本计画将联合收割机结合全内反射荧光（TIRF）与光漂白后荧光再分布（FRAP）两种成像模式于一台显微镜上。具体而言，现有的多激光TIRF显微镜将被改造为必要的硬件，使FRAP能够进行，并生成关于细胞表面蛋白质动态周转的数据。这种新工具对于量化合成世界上最丰富的生物聚合物纤维素的酶复合物以及位于质膜上的膜受体、转运蛋白和其他蛋白质复合物的营业额将是不可或缺的。