The VA Tennessee Valley Healthcare System (VATVHS), Nashville campus is requesting funds to purchase a Zeiss LSM980 Airyscan confocal microscope. The LSM980 is a state-of-the-art point-scanning confocal microscope with particular functionality for super-resolution, live cell, and large mosaic imaging. The LSM980 will be available for VA investigators and maintained by the Vanderbilt Cell Imaging Shared Resource (CISR). The addition of this new instrument is crucial to support the ongoing research of eight VA investigators represented as Major Users Projects in this proposal. These investigators all have active VA funding and essential needs to advance research topics including diabetes, C. difficile infection, Inflammatory Bowel Disease (IBD), extracellular matrix and fibrosis, chronic obstructive pulmonary disease (COPD), kidney development, and gastric cancer. We expect that numerous other research programs at the VA and within the wider Vanderbilt community will also benefit from this instrument; particularly those that require super- resolution, live sample, and large tissue imaging. This Zeiss LSM980 Airyscan will offer dedicated confocal hours for these VA researchers as well as greater resolution afforded by Airyscan and increased super- resolution imaging speeds. We chose the LSM980 as a significant need for these 8 VA users based on three main goals: 1) to provide additional confocal hours because existing CISR confocal microscopes are saturated, 2) to add a confocal microscope dedicated to VA investigators who need longer imaging sessions for large samples and for live cell imaging, 3) to offer new capabilities including the latest generation Airyscan for 4x faster super-resolution imaging and state-of-the-art detectors for increased sensitivity. The requested instrument is configured based on the needs of the Major Users in consultation with PI Dr. Roy Zent and Cell Imaging Shared Resource personnel. The full system includes the LSM980 confocal with state-of-the-art GaAsP PMTs, Airyscan super-resolution module, incubation and environmental control, a variety of objective lenses, 6 excitation lasers, and an acquisition workstation with ZEN 3.1 software. Significant consideration was given to optical performance in choosing the variety of objective lenses to accommodate samples including fixed or live cells, fixed or live tissue, and large optically cleared tissue. The latest generation of detectors offers greater sensitivity for signal detection and gentle imaging for live cell experiments. The new Airyscan2 module offers a separate detector to achieve resolution up to 1.7x greater than the limit of conventional confocal microscopy. The environmental chamber equipped with 37 C incubation, 5% CO2, and thermal drift correction allows live cell imaging from minutes to hours to overnight. Excitation lasers including 405, 445, 488, 514, 561, and 639 nm are appropriate for a wide range of modern fluorescent proteins and dyes. These LSM980 accessories are fundamental to the successful implementation of the advanced imaging described in the Major User projects. The expertise and dedicated support for this instrument are exceptional. The CISR has a 20+ year history of assisting over 400 investigator labs with cutting-edge imaging expertise and cooperative research support. PI Dr. Roy Zent is a Staff Physician and VA investigator with expertise in kidney development and high-resolution microscopy and is Major User on this project. Additionally, all 8 Major Users in this proposal are VA investigators with confocal microscopy experience.
Confocal microscopy is one of the most prevalent techniques for imaging of cellular anatomy and physiology. The Zeiss LSM980 Airyscan confocal microscope will support VA investigator research in diabetes, C. difficile infection, Inflammatory Bowel Disease (IBD), extracellular matrix and fibrosis, chronic obstructive pulmonary disease (COPD), kidney development, gastric cancer, and more. There is a substantial and fundamental need for this state-of-the-art instrument to support the ongoing work of numerous VA-sponsored investigators making vital contributions to health care discovery and innovation within the veteran population.