Super-resolution confocal microscope

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ABSTRACT The La Jolla Institute for Allergy and Immunology (LJI) is a premier immunology research institute in the US whose 24 principal investigators are supported by ~$40 million in NIH grants and contracts. At LJI we study the behavior of immune cells in diseases prominent in the US and around the world, including: type 1 diabetes, atherosclerosis, rheumatoid arthritis, and Zika infection. Nearly all of our scientists use laser scanning confocal imaging to visualize cellular interactions, signaling, and function and the results have been published in Nature, Science, and other prominent journals. However, to progress further in this important work, we need a versatile confocal microscope with super-resolution capabilities. Currently, LJI lacks a dynamic, capable, and technologically current confocal system and needs one that will yield the most impactful data for each NIH research project. This need includes a host of fluorescence based assays such as quantitative receptor translocation, protein co-localization intracellularly and in tissue, fluorescence resonance energy transfer (FRET), and fluorescence recovery after photobleaching (FRAP), and photoactivation. We have a 10 year old Leica SP5 combined confocal and multiphoton system that is dedicated to intravital imaging and a simple automated Olympus FV10i confocal suitable for fixed cells and tissues at lower resolutions. LJI has no super-resolution capability. Here, we seek support for purchasing a state-of-the-art Zeiss 880 confocal microscope with an Airyscan super-resolution detector, highly sensitive detection devices (GaAsP detectors), the greatest signal-to-noise ratio system for both fixed and live acquisition, multiple laser lines covering the entire visible light spectrum, and a fully tunable spectral emission collection grating. Airyscan is a novel super-resolution technology that can achieve 140 nm resolution using standard immunofluorescence protocols. After comparing all instruments on the market, it is clear that the Zeiss LSM 880 system is best suited for the needs of LJI investigators and, if funded, it will undoubtedly have a significant impact on NIH-supported scientific progress.