MRI Development of a fast P3D-STED Microscope

University Of Colorado At Denver-Downtown Campus, Denver CO

Investigators

Abstract

Title: MRI Development of a FAST P3D-STED Microscope An NSF MRI Development Grant award is made to a collaborative team of physicists, engineers, and neuroscientists at the University of Colorado Denver to further develop the capabilities of Stimulated Emission Depletion (STED) microscopy. The goal of the project is to develop a FAST P3D-STED microscope (fast acquisition, photoactivation, 3-dimensional enhanced resolution STED microscope). Historically, light microscopy has been limited in its resolution by the Diffraction Barrier - a fundamental physical limit due to the property of photons of spreading out when they pass by an edge, such as the aperture in a microscope. In the past decade, several methods have emerged that can "break" the diffraction barrier, causing a revolution in the field of biological imaging and opening a major window of opportunity for studies that heretofore have been impossible. STimulated Emission Depletion (STED) microscopy, invented by Stefan Hell, is one such super-resolution technique, allowing fluorescence imaging at resolutions of tens of nanometers. With continuing improvements in the technology, STED is now able to be utilized with a wide range of fluorescent dyes and genetically encoded fluorophores and has advantages for fast, live cell, and in vivo imaging. The investigators will further push the current capabilities of STED microscopy by developing a new STED microscope to allow high speed, sub-diffraction limit imaging combined with photoactivation capabilities. This microscope will allow researchers to not only observe small objects with sub-diffraction resolution, but also enable control of dynamic processes and observation in real time. The projects enabled by the instrument include studies of the dynamic organization of protein complexes in synapses upon control of neural plasticity, studies of the molecular-level mechanisms of odor transduction by direct stimulation of the transduction pathway, and methods for writing/reading bits at sub-diffraction dimensions in high density data storage materials. The potential societal impacts of this research include improved treatment of neurological disorders and advances in information handling and storage. The FAST P3D STED microscope will be housed in the Advanced Light Microscopy Core (ALMC) facility at the University of Colorado Denver, a successful shared user facility that has personnel and resources to train new users and operate and maintain the instrument. Researchers, postdoctoral trainees, and students from a variety of science and engineering departments at the University of Colorado Denver, University of Colorado Boulder and other universities will be able to utilize the microscope for their research. Outreach to new users will be accomplished through seminars put on by the ALMC and research presentations at scientific meetings. The investigators are particularly committed to promoting student training and advancing participation of under-represented minorities in science as evidenced by their involvement in the Building Research Achievement in Neuroscience (BRAiN) program, a summer program designed to help meet the challenge to reduce the Neuroscience research participation gap by preparing diverse undergraduates in the Rocky Mountain and Southwest Region for successful entry to Neuroscience Ph.D. programs. The capability of the STED microscope to see the inner workings of cells at unprecedented resolution captures the imagination of these future scientists.