This Major Research Instrumentation award funds the acquisition of a multi-photon laser scanning confocal microscope for research and training in interdisciplinary fields on the University of Oklahoma (OU) Norman campus. The new instrument has higher sensitivity and faster speed and will significantly boost the capabilities for live and deep-tissue imaging by a diversity of life science and engineering researchers on the OU campus and at nearby research institutions. This new confocal microscope will be used by a number of NSF-sponsored laboratories that are already active in cell and molecular imaging as well as by new users who will incorporate confocal imaging in their research projects. The research topics include, but they are not limited to, neural crest migration in the primitive vertebrate lamprey, neurite outgrowth and synaptic vesicle trafficking in fruit flies, calcium imaging of deep nerve tissues, temporal and spatial patterns of protein complex formation in plants, and biofilm growth and morphology during corrosion of metallic surface. The new microscope will be an important part of education and research integration; students will be trained initially through a new upper undergraduate and graduate student course, Applied Confocal Imaging, taught collectively by the PIs. In addition, the PIs and their students will collaborate with rural and urban Oklahoma K-12 teachers in developing and implementing cutting-edge outreach and classroom projects. The results of these research, teaching, and outreach efforts will be broadly disseminated through participation of students and faculty at professional meetings and through peer-reviewed publications.
This project established a light microscopy core facility on the University of Oklahoma-Norman campus. The facility houses a Leica SP8 confocal microscope that was installed in December 2012. This instrument is equipped with a Coherent Ti:Sapphire laser for multiphoton (live deep-tissue) imaging, and FLIM capability for fluorescence lifetime imaging. Since its establishment, this core facility has grown to house an Olympus FV500 and a Zeiss Axioimager Apotome that are used for routine fluorescence imaging. Beginning in January 2013, the SP8 has logged approximately 1000 hours of usage, generating data that have been included in 7 manuscripts either submitted or already published, one book chapter, and four conference presentations. The instrument has been used by 17 different research groups from 5 departments and includes users both on and outside the OU-Norman campus. Intellectual Merit: Modern imaging is indispensible in today’s biological research, yet prior to this project, the only confocal microscopes available at the University of Oklahoma were ~10 years old and functionally limited. The instrument purchased in this project has significantly boosted the capabilities for live and deep-tissue imaging by a diversity of life science and engineering researchers on the OU campus and at nearby research institutions. The intellectual merits of the scientific experiments enabled by this project have been significant in several aspects, and reflect the broad capabilites of the instrument for a diverse user group. For example, using multi-photon capabilities, researchers have performed deep-tissue imaging with live preparations. This has enabled them to study cell lineage analysis in the primitive vertebrate lamprey and to observe neurite outgrowth in the fruit fly Drosophila. Such studies may shed light on the evolutionary mechanisms underlying vertebrate development and on neuronal remodeling during insect metamorphosis. Second, the new system has enabled tracing of synaptic vesicle recycling in Drosophila to provide novel insights into the dynamic process of neuronal function at the single synapse and neural circuitry levels. Third, the system has allowed for imaging of large electrically excitable cells in weakly electric fish. This work provides a foundation for the development of biogenerators to power implantable medical devices. Fourth has been the ability to perform Förster resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM) experiments to facilitate the examination of temporal and spatial patterns of protein complex formation in plants. Broader Impacts: This project has had significant broader impacts on education, integration of research, public outreach, and student mentorship. Specifically, the microscope and core facility have been integral to a microscopy course that was developed as a part of this project that has resulted in the development and dissemination of new educational materials. To date, this course has trained over 20 undergraduate and graduate students in research that uses fluorescence microscopy tools. Independent research by more than 15 post-doc, graduate, and undergraduate students has also integrated the use of confocal and multiphoton microscopy as a part of STEM training. In addition, this project has impacted K-12 STEM education in Oklahoma. Science teachers from two area high schools participated in a summer research project in 2013 that used the SP8 microscope. Their experience has impacted over 150 high school students to become engaged in understanding the importance of biofilms in everyday life. These teachers were supported by the grant "RET Site: Strengthening a K12 Learning Community through Engineering Research", leveraging both of these projects for maximum benefit to students in science education.
