Super-resolution confocal microscope - 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.
