Cell & Molecular Redox Imaging Core – Project Summary The South Carolina COBRE in Oxidants, Redox Balance and Stress Signaling (Redox COBRE) Cell and Molecular Imaging Redox Core (CMRI) provides advanced technologies and expertise (Aim 1), capabilities for new method development (Aim 2), and training (Aim 3) to target investigators and Center members for state-of- the-art cell- and tissue-based microscopic imaging. This includes confocal, multiphoton and super-resolution microscopy of live and fixed cells and tissues, intravital imaging, and automated imaging of histological slides. Additionally, the Core maintains and develops a variety of molecular tools and redox indicators for use in COBRE investigations. The CMRI houses the following major microscope systems: 1) Zeiss LSM 880 NLO multiphoton/confocal system equipped with a Coherent Chameleon multi-photon laser, Quasar spectral detection and Airyscan super-resolution capability; 2) Olympus Fluoview FV1200 multiphoton microscope with SpectraPhysics MaiTai DeepSee laser and silicone oil optics for intravital imaging; 3) Olympus Fluoview FV 10i LIV live cell confocal microscope with water immersion optics; 4) Zeiss LSM 510 META confocal microscope; 5) BD BioSciences CARV II disk-scanning confocal microscope for video-rate imaging; 6) Zeiss Axiovert 200M wide-field fluorescence microscope; and 7) Perkin-Elmer Vectra Polaris Automated Quantitative Pathology Imaging System. Except for the Vectra Polaris, which is customized for histological slides, all microscopes are equipped with environmental chambers for temperature and gas phase control to allow non-destructive 3D imaging of living cells, tissues and organisms. Major applications include: 1) live cell imaging of parameter- sensitive fluorophores to monitor ions, electrical potentials, radical generation, pyridine nucleotide reduction, membrane permeability, cell viability (apoptosis and necrosis), and the submicron distribution of fluorescent proteins and other fluorescent reporters; 2) high resolution imaging of tissue sections for immunocytochemistry and fluorescent protein distribution; 3) fluorescence resonance energy transfer (FRET) and DuoLink to characterize and quantify interactions between specific molecules; 4) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization and permeability, radical generation, gene expression and other parameters in living animals; and 5) high throughput, quantitative multiplexed imaging of conventionally and immunostained clinical and research specimens. Ancillary equipment required for specimen preparation is also provided. Consultation and services for transmission and scanning electron microscopy (TEM and SEM, respectively) are available through the Department of Pathology & Laboratory Medicine. Computer workstations provide offline image processing/analysis (ImageJ FIJI, Metamorph, IPLab and other software).