Acquisition of a Zeiss LSM 900 confocal microscope with Airyscan 2 for an Imaging and Microscopy Core

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Research Summary This grant proposal seeks funding for a Zeiss LSM 900 laser scanning confocal microscope with Airyscan 2 for NIH funded investigators who use the Advanced Imaging and Microscopy Core Facility at Loma Linda University to perform both histochemical analyses and real-time imaging of cells and tissues. On the one hand, the system would be used to localize various proteins and cells using fluorescent labeling approaches, while on the other it would be used to examine the activity of living cells and tissues. These two imaging technologies are required as the NIH funded major and minor user group has diverse needs. Their research encompasses cardiovascular physiology, pregnancy, development, neurobiology, neuroendocrinology, cancer biology, radiation medicine, regenerative medicine, and health disparities, but all have common interests in measuring the spatial apposition between cells, changes in protein expression, localization of proteins within cells, and spatial and temporal sequences involved with cell signaling. The research record of the user group is excellent, representing some of the most successful investigators at the institution. The Zeiss LSM 900 confocal would support numerous NIH research grants and investigative programs across campus and training programs. The core facility has two shared-use confocal microscopes that are near or at their end of functional use. Acquisition of a Zeiss LSM 900 with a AiryScan 2 will solve this pressing issue and will enable funded researchers to continue their studies including cell signaling, in-situ hybridization, protein expression and localization, cell and tissue morphology, interactions between molecules and proteins, spatial reconstruction, spatial-temporal relationships, and subcellular transport. These questions require the ability to image single or multiple fluorescent probes in either fixed or live preparations. Based on the needs of the funded projects, a Zeiss LSM 900 confocal microscope is an ideal replacement instrument. The instrument has 2 high-sensitivity confocal channels for recording fluorescence; one for transmitted light (DIC) and has an Airyscan 2 unit for super-resolution imaging along with adaptive focusing capabilities and can perform mosaic tiling for multidimensional reconstruction. The Airyscan 2 will also allow for rapid confocal imaging for spatial and temporal recordings of cell signaling systems, which are key aspects to multiple awards. The Zeiss LSM 900 offers a fully integrated system available for the wide breadth of studies of the user group and will provide continuity with an existing multiphoton Zeiss 710 system. The institution recognizes the quality of the facility and user group and has strongly supported the facility for over a decade with full funding for staffing, supplies, and service contracts. The PI has considerable experience with confocal microscopy and will oversee the daily operation of the instrument and be part of a multidisciplinary internal advisory committee that oversees the core facility. The instrument will be housed in the existing AIM facility, providing continued contemporary instrumentation for this highly used shared-use imaging core.