Fluorescence Microscope for Automated Large Array Scanning and 3D imaging

Colorado State University, Fort Collins CO

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Abstract

Project Summary This proposal seeks funding for a Keyence BZ-X710 all-in-one fluorescence microscope, which provides a wide range of wide-field imaging capabilities in concert with incredible ease-of-use. The unique set of microscopy capabilities provided in this microscope permit high throughput, rapid imaging of a vast array of fixed specimens in various preparations, e.g., multi-well microtiter plates, culture dishes, microscope slides, roller tubes, etc. Further, the BZ-X710 incorporates an optional stage-top incubation chamber to enable imaging of live cells and in vitro tissues. Another advantage of the BZ-X710 that has a transformative impact on live cell imaging is the high speed at which large images can be acquired and stitched into a mosaic, with demonstrated repeatability to well below the spatial resolution of the imaging system. Finally, the nature of the illumination system significantly decreases deleterious effects such as photobleaching and photodamage, and thus both living and fixed specimens can be imaged for longer periods of time than with, for example, a confocal microscope. The structured illumination system and deconvolution algorithms allow for rapid imaging of relatively thick tissue slices (up to 500 ?m) at multiple wavelengths. This combination of unique attributes permits the use of BZ-X710 for long-term fluorescent imaging studies of living cells and tissue slices. Observation of living cells with a large number of exposures is paramount for studying long-term behavior of a specimen, and could greatly aid in elucidating the function of biomolecules within the cell. Here we will exploit the power of the BZ-X710 by applying it to many projects of biomedical significance. These projects ask questions whose answers will improve our understanding of basic cell biological phenomena and apply this knowledge to diverse areas of human health, such as neurodegenerative disorders and cancer. The ease-of-use of the Keyence BZ-X710 microscope allows us to incorporate this technology into a microscopy core facility that will enable its use by many research groups, while maintaining access to the major users who have proposed many exciting questions to be addressed here. Incorporation into a Foundational Core and having oversight, training and data acquisition and analysis provided by a university funded facility director with an applied physics PhD and several years of experience in bioscience applications will ensure proper long-term operation.