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

这项合同将使购买MINFLUX成像和分子跟踪系统成为可能。MINFLUX技术可以定位到2纳米分辨率的分子，并能够以10 MHz的时间分辨率跟踪分子，这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斯特凡·地狱开发的一项技术。顾名思义，MINFLUX通过照亮周围的分子而不是将分子本身暴露在高光强度下来检测单个分子，大大减少了漂白和光毒性。这是通过光学激活荧光分子，然后用环状激发光包围它来实现的。当外环的中心从分子上移开时，荧光强度增加，表明分子的位置。然后，这个过程在三个方向上进行，在空间中定位分子，同时将分子经历的光子流量减少到每个定位事件800到2500个光子的范围。这个仪器的非凡能力将允许来自伊利诺伊大学10个参与单位的至少20名教职员工跟踪神经元突触、图像染色质折叠、神经肌肉连接等中的受体。都是在细胞或其他复杂环境的背景下进行的。目前，美国有两个MINFLUX系统。一个位于西海岸的斯克里普斯，一个位于东海岸的NIH。该地区的MINFLUX仪器将为位于中西部的教职员工提供途径，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开展新的科学，并促进实验生物物理学家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，通过相同的培训程序、日历和账单进行管理。作为MINFLUX在中西部的早期采用者，IGB基金将与Abberior Instruments合作，为校外研究人员提供演示和访问机会，并为伊利诺伊州和整个中西部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优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.