MRI: Development of a scan-less temporal focusing two-photon fluorescence microscope for high-speed three-dimensional imaging

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An award is made to the University of Texas at El Paso (UTEP) to develop a high-speed optical microscope for imaging fluorescent objects and simultaneously tracking their fast motion in three-dimensional (3D) space. This interdisciplinary project enriches undergraduate and graduate students' education by providing frontier research experience in the fields of biology, chemistry, physics, and electrical engineering. It also trains postdoctoral researchers who will form the next generation of instrumentalists. This project gives scientists in biology and chemistry a strong background in the development and use of innovative and contemporaryinstruments, significantly enhancing their ability to conduct cutting-edge research. The developed microscope will serve as the cornerstone of a Biophotonics core facility that will promote collaborative research, education and outreach activities. Underrepresented minority students at UTEP, a Hispanic-majority (almost 80%) institution, and regional users will be invited to participate in workshops and research activities using this facility.This project is an innovation in the field of fluorescence microscopy. By integrating two ultrafast laser techniques, temporal focusing and pulse shaping, this new microscope has the capability of acquiring 3D volumetric images at an unprecedented high-speed (50 volumes/s, volume size 100um × 100um × 100um). This microscope eliminates the current laser beam scanning mechanisms typically used in confocal and two-photon microscopes with temporal focusing, and adds depth scanning capability by integrating the pulse shaping technique. This project will also develop a computerized image processing system that will automatically analyze high-speed videos. These capabilities, i.e. high-speed and 3D volumetric imaging, will enable researchers to explore and analyze previously unobtainable fast dynamic information from this automatic image analysis systems. Initial applications are in the areas of marine virus infection, nanoparticle transportation, and fly brain imaging. Applications could extend to many other fields where it is hard to study fast 3D dynamic processes using current laser beam scanning fluorescence microscopes.