The NSF-Major Research Instrumentation program will enable Trinity University of San Antonio, TX to acquire a Nikon A1 confocal microscopy system. Confocal microscopy technology produces high resolution, three-dimensional, multicolor images or videos obtained from live or fixed biological specimens. Biological systems are dynamic and regulated through complex interactions between molecules. The confocal microscope technology optically sections through the sample, minimizing out of focus light to yield superior image quality and three-dimensional image reconstructions. The confocal microscope will allow investigators to determine spatial and temporal interactions through the use of multi-color fluorescence labeling. Research by the PI and four coPIs addresses a diverse array of biological problems, including epithelial cell junction homeostasis, metabolism and autophagy in the central nervous system, segmental regeneration, microbial metabolite biosynthesis, oxidant signaling and mitochondrial function, muscle fiber type analysis and mathematical modeling of cell migration. High-speed, multicolor, three-dimensional reconstruction with submicron resolution capability permits discovery and analysis of complex biological phenomena in all of these areas. The confocal system equipped with the spectral detector minimizes background signal and allows investigators to use dyes with overlapping spectral characteristics. Colocalization and molecular studies require the high-speed resonant scan-head and spectral detector for accurate, real-time spectral differentiation to resolve transient interactions. The environmental chamber in combination with the Perfect Focus system enables live cell research of sub-cellular components and imaging of molecular interactions in real-time.
The Nikon A1 confocal microscope system is to be housed in the Imaging Suite within the new Center for Sciences and Innovation at Trinity University. The Imaging Suite will bring together Trinity University faculty and student researchers as well as faculty and student researchers from local institutions, including the University of the Incarnate Word and St. Phillips Community College, to address a range a biological questions requiring confocal microscopy. The investigators have an established record for engaging undergraduate students in their research and using their research programs to develop innovative learning experiences for the classroom and teaching laboratory. Undergraduate science courses in Biology, Neuroscience, Mathematics and Physics will experience how confocal microscopy may be used in discovery and investigations. Trinity University will host a series of confocal microscopy workshops for the area that address both theoretical and practical considerations for this technology. The investigators are also partnering with local science educators and undergraduate students planning careers in science education.
was support by the National Science Foundation-Major Research Instrumentation (NSF-MRI) award to Trinity University. A group of five faculty members from two neighboring institutions in San Antonio (Trinity University and University of the Incarnate Word) have been interacting to enhance the research and learning capability at our given institutions. Faculty and students have had opportunity for multiple in-depth training sessions that were tailored specifically to particular research questions. These training opportunities provided investigators to learn new methods of data collection and analysis while considering alternate approaches to research questions. Sophistication of approach has increased as research teams gained expertise on the confocal microscope. Conservatively, more than two-dozen undergraduate researchers have had opportunity to participate and conduct studies in a collaborative nature with the project PIs. Numerous scientific products have been generated as a result of these interactions including talks, presentations at scientific meetings and peer-reviewed publications. Intellectual Merit: Through faculty-driven research questions spanning a diverse array of biological problems have been investigated using the confocal microscope including epithelial cell junction homeostasis, segmental regeneration, lipid metabolism, mitochondrial function and bacterial chemotaxis. Importantly, many of these project have been multidisciplinary teams that have engaged undergraduate student in the process of science. Dr. King’s research has employed a molecular structure-function approach to examine the dynamics of ZO-1 a cell junction scaffold. Cell junction formation is essential for multicellular life and loss of cell junctions is a hallmark of numerous disease processes. Transgenic cell lines were constructed that express fluorescent protein-ZO-1 mutants which were then used for fluorescent recovery after photobleaching (FRAP) experiments. These experiments demonstrate the importance of specific ZO-1 domains in providing stabilizing interactions at the cell junction. Dr. Healy’s project has developed integrated population-level bacterial chemotaxis models incorporating the fluid environment with physiological and biochemical parameters. Using applied mathematics and fluid dynamics approaches, 2D and 3D chemotaxis models were generated for chemically heterogeneous fluid environments. Laser scanning confocal microscopy was used to observe and analyze 2D bacterial swimming behavior. These models are beginning to yield valuable information on how bacterial swimming behavior is affected by fluid dynamic properties of their natural environments. Dr. Acosta’s project has characterized the expression of the developmental marker, beta catenin, in the nervous system of a regenerating model system, Lumbriculus variegatus. Confocal analysis highlights beta-catenin expression in the ventral nerve cord with visible up regulation in the lateral giant fiber and in smaller intermediate giant fibers which run the length of the nerve cord extending connections which cross the ventral nerve cord at segmental boundaries. We continue the use of this novel model system to further develop a better understanding of the role of beta-catenin and other molecular mechanisms involved in nerve regeneration and wound healing. Broader Impacts: The acquisition of the confocal microscope has impacted teaching climate as practical experiences have transitioned to inquiry-based investigations. Development of curricular elements in Cell Biology, Neurobiology and Physiology provide students access to research grade microscopy to address contemporary issues within the disciplines. These pedagogical approaches lower the threshold for students moving into intensive research experiences and into innovative scientific enterprises. A meaningful and motivated population of under-represented individuals in science have access to the confocal microscope through teaching and research programs. On-campus high school groups (e.g. Duke TIPS) students met with faculty and students using the confocal while discussing scientific interests and career aspirations. The VIEW symposium hosted by the principal investigators was a large-scale event to deepen interest in various modes of scientific imaging. Speakers from a variety of disciplines and student presenters demonstrated how scientific imaging is used to address a broad array of problems. The programs described that have been launched in coordination with the acquisition of the confocal microscope are anticipated to continue beyond the funding interval.
