Acquisition of Two-Color 3-D Minflux for Live-Cell Single Molecule Imaging and Tracking at Unprecedented Spatial and Temporal Resolution

采集双色 3-D Minflux，以前所未有的空间和时间分辨率进行活细胞单分子成像和跟踪

• 批准号: 2241074
• 负责人: Martin Gruebele
• 金额: $ 19.45万
• 依托单位: University of Illinois at Urbana-Champaign
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2241074&HistoricalAwards=false
• 关键词: Acquisition; Two; Color; Minflux; Live

This award will enable the purchase of a MINFLUX imaging and molecule tracking system. MINFLUX technology can locate molecules to 2 nm resolution and the ability to track molecules with 10 MHz time resolution an order of magnitude improvement over other single molecule techniques. MINFLUX is a technology developed by Nobelist, Stefan Hell. As the name suggests, MINFLUX detects individual molecules by illuminating their surroundings and not exposing the molecule itself to high light intensities, greatly reducing bleaching and phototoxicity. This is accomplished by optically activating a fluorescent molecule and then surrounding it with a donut of excitation light. As the center of the donout is moved off the molecule the fluorescence intensity increases indicating the position of the molecule. This process is then carried out in three directions locating the molecule in space while minimizing the photon flux experienced by the molecule to the range of 800 to 2500 photons per localization event.The extraordinary capabilities of this instrument will allow at least 20 faculty from 10 participating units at the University of Illinois to track receptors in neuronal synapses, image chromatin folding, neural muscular junctions, and much more. All against the background of the cell or other complex environments. There are currently two MINFLUX systems in the United States. One is located at Scripps on the west coast and one at NIH on the east coast. A MINFLUX instrument in the area would provide access to faculty members located in the Midwest and strengthen the University of Illinois’ standing in imaging.The best-in-class resolution of the MINFLUX microscope will allow new science to be conducted and foster collaborations between experimental and theoretical biophysicists to characterize these discoveries. It will be housed in the IGB Core Facility and will be managed with the same training procedures, calendars, and billing, as our other high end optical microscopes. As the MINFLUX early adopter in the Midwest, the IGB Facility will work with Abberior instruments to provide demonstrations and access for off campus researchers along with hands- on training for students and postdocs at Illinois and throughout the MidwestThis award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该合同将使MINFLUX成像和分子跟踪系统的采购成为可能。MINFLUX技术可以定位到2nm分辨率的分子，并且能够以10mhz的时间分辨率跟踪分子，比其他单分子技术提高了一个数量级。MINFLUX是由诺贝尔奖获得者Stefan Hell开发的一项技术。顾名思义，MINFLUX通过照亮周围环境来检测单个分子，而不会将分子本身暴露在强光下，从而大大减少了漂白和光毒性。这是通过光学激活一个荧光分子，然后用一个环形的激发光包围它来实现的。当圆环的中心被移出分子时，荧光强度增加，表明分子的位置。然后在三个方向上进行这个过程，在空间中定位分子，同时将分子所经历的光子通量最小化到每个定位事件的800到2500个光子。该仪器的非凡功能将允许来自伊利诺伊大学10个参与单位的至少20名教师跟踪神经元突触中的受体，图像染色质折叠，神经肌肉连接等。所有这些都是在细胞或其他复杂环境的背景下进行的。目前在美国有两个MINFLUX系统。一个在西海岸的斯克里普斯另一个在东海岸的国家卫生研究院。该地区的MINFLUX仪器将为位于中西部的教职员工提供服务，并加强伊利诺伊大学在成像方面的地位。MINFLUX显微镜的最佳分辨率将允许进行新的科学研究，并促进实验和理论生物物理学家之间的合作，以表征这些发现。它将被安置在IGB核心设施中，并将与我们的其他高端光学显微镜一样，采用相同的培训程序，日历和计费管理。作为中西部地区MINFLUX的早期采用者，IGB设施将与Abberior仪器合作，为校外研究人员提供演示和访问，并为伊利诺伊州和整个中西部地区的学生和博士后提供实践培训。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。