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

University Of Illinois At Urbana-Champaign, Urbana IL

Investigators

Abstract

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 Midwest This 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.