With this award from the Major Research Instrumentation (MRI) program, Drs. Watson, LaRiviere, Stewart, Erickson, Turner and ten colleagues from the Departments of Biology, Psychology, Physics and Engineering, and Computer Science, along with the Programs in Neuroscience, Biochemistry, and Environmental Sciences at Washington and Lee University (W&L), Virginia Military Institute (VMI) and Mary Baldwin College (MBC) will acquire an Olympus Fluoview 1000 (FV1000) Spectral Confocal Live Cell System for research and training across the sciences. This confocal laser scanning microscope (CLSM) will form the foundation of the newly created communal microscopy, imaging, and computational core of a projected Interdisciplinary Quantitative Science Center that is part of a long-range, grant-funded initiative at W&L which promotes mathematical modeling, bioinformatics, and data analysis. The confocal microscope will enable fifteen researchers from three schools to expand their ability to detect, quantify, and localize gene products and to study biological structures, thereby expanding the scope of existing research agendas and developing new as well as collaborative research opportunities. Specifically, the confocal microscope will facilitate and impact the following areas: 1) the study of different aspects of synaptic connectivity and influences on neuronal wiring in the brain and central nervous system, 2) eukaryotic nonfunctional ribosomal RNA decay pathways, 3) effects of hormone regulation in the developing cardiovascular system and sexual differentiation, 4) mechanism of Ca++ regulation, 5) reconstruction of glandular morphology of terrestrial invertebrates, 6) ultrastructural analysis of single-celled eukaryotes, 7) rapid assessment of otolith structure in new fish species, 8) comparison of the formation of dense core secretory granules cells, 9) localization of nitrogen-cycling microorganisms on freshly harvested fine roots and organic matter and 10) image segmentation and analysis studies.

Confocal microscopy is a technique that allows a three dimensional high resolution image acquisition of live or fixed specimens. By attaching fluorescent dyes (fluorophores) to biological specimens, cells and sub-cellular components can be identified with a high degree of specificity amid non-fluorescing material. Moreover, several target molecules can be visualized simultaneously with multiple fluorophores emitting light at differing wavelengths. Using image analysis software, the acquired serial optical sections are then rendered to generate a clean high resolution three-dimensional reconstruction of the specimen in which all out-of-focus light has been rejected. The result is an exceptionally clean high resolution image of a biological specimen that can reveal the presence of a single molecule. This instrument is critical for the study of biological structures and will have a transformative effect on how emergent young scientists are trained at W&L, VMI, and MBC. The CLSM will provide cutting edge tools for faculty to carry out their research projects and provide microscopy training and research opportunities to undergraduate students from three rural primarily undergraduate institutions. Using the teacher-scholar model, which seamlessly integrates research with discovery-based laboratory course work, W&L and VMI, will take full advantage of this instrumentation to promote further research integration into courses, to expand undergraduate programmatic research opportunities, and to promote student and faculty collaborations within and between departments and institutions.

Project Report

This NSF-DBI Award (1126118) supplied funds to acquire a laser scanning confocal microscope (LSCM) to expand research and training across the sciences at two primarily undergraduate institutes (PUIs): Washington & Lee University (W&L) and neighboring Virginia Military Institute (VMI). This LSCM provides a critical piece of high-end instrumentation that supports the hands-on science pedagogy of liberal arts colleges, which prepare a disproportionately large number of students for careers in Science, Technology, Engineering, and Math (STEM) fields. Although housed at W&L, the LSCM is equally accessible to VMI as both campuses are contiguous. Intellectual merit – The LSCM allows users to study intricate biological structures and to detect, quantify, and localize gene products in numerous biological systems. Thus, the LSCM has greatly expanded the scope of existing research programs at W&L and VMI, and it has led to new and collaborative research opportunities at these institutions. Faculty and students from W&L and VMI have used the LSCM to address questions that span basic molecular processes to systems level structure and function. Seven faculty members have used the LSCM to expand their research and training activities with undergraduates. For example, W&L biology professors Fiona Watson and Sarah Blythe, and W&L psychology professor Robert Stewart use the confocal microscope to study synaptic connectivity and influences on neuronal wiring in the brain. Specifically, Watson uses the LSCM to explore the regenerative capacity of retinal ganglion cells (RGCs) in the eye of the South African frog (Xenopus laevis) and has initiated an entirely new research project exploring the effects of organophosphate insecticides on the cytoarchitecture of developing spinal sensory neurons. Blythe uses the LSCM to examine the neuroanatomical changes associated with diet-induced cognitive impairment in male and female rats. Stewart uses the LSCM to study postnatal changes in taste primary afferent terminations in the first central taste relay nucleus. Four other faculty members use the LSCM for various and diverse biological applications. For example, W&L chemistry/biochemistry professor Fred LaRiviere investigates targeted degradation of functionally defective ribosomes in Saccharomyces cerevisiae. This is a collaborative project with W&L biology professor Gregg Whitworth who also uses the LCSM to quantify the real-time, in vivo, rate of apoptotic induction in S. cerevisiae under a variety of environmental stress conditions. VMI biology professor James Turner has used the LSCM to view the developing vascular system of the transgenic zebrafish (Allgood et al., 2012). Finally, the LSCM helped recruit new W&L chemistry/biochemistry professor Kyle Friend who is using the LSCM to image RNA movement and cell division patterns in frog embryos. Friend also began a research collaboration with Watson to investigate the mechanism of inter-cellular RNA transfer. Research projects with the LSCM have resulted in two peer-reviewed publications and five national or international conference abstracts. Finally, the LSCM was used to produce preliminary results needed for submission of two basic research funding grants (pending). Broader impacts – The LSCM is housed in the new Integrative and Quantitative (IQ) Center at W&L. This is a unique, centrally-located, space that is home to sophisticated imaging and computational instruments, including a scanning electron microscope, and a powerful 3-D imaging lab. The IQ Center serves to foster cross-disciplinary and quantitative approaches to inspire creativity and problem solving in teaching and research (see: www.wlu.edu/iq-center). The LSCM in combination with the other instrumentation is having a transformative effect on how students are trained. In fact, since the space opened in September 2013, the IQ Center has been used by 52 faculty members from 25 departments across the undergraduate campus in teaching 42 courses. Thus, 926 students from all STEM disciplines, as well as from departments and programs as varied as English, Latin, Accounting, History, Art and Dance have been exposed to the space and its technologies in their classes. At W&L and VMI, the teacher-scholar model is used to seamlessly integrate research with discovery-based laboratory course work. Thus, we use the LSCM to promote further research integration into courses and to expand undergraduate programmatic research opportunities. Already, the LSCM has been used to train undergraduates in laboratory components of three upper level biology and neuroscience courses and has been used by numerous students for their research projects with W&L and VMI faculty. Indeed, approximately 50 undergraduate students have used the LSCM during their undergraduate experience, three of which have co-authored two peer reviewed publications and six have presented at international conferences. Given the centrally-located space and use by faculty in research and courses, the confocal will continue to receive wide exposure and help recruit competitive faculty and students well beyond the final grant award period. Indeed, six additional faculty have already outlined plans to expand their research using the LSCM and two other faculty have plans to introduce the LSCM into their course curricula.

National Science Foundation (NSF)
Division of Biological Infrastructure (DBI)
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Program Officer
Robert Fleischmann
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Washington and Lee University
United States
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