CELL & MOLECULAR IMAGING (CMI) SHARED RESOURCE: SUMMARY The Aims of the Cell & Molecular Imaging (CMI) Shared Resource are to: 1) provide HCC members with technologies, expertise, and training for state-of-the-art confocal, multiphoton, and super-resolution microscopy, including image processing and analysis; 2) develop novel imaging methods and applications to meet emerging scientific needs; and 3) educate, train, and consult on cutting-edge technologies. Microscope systems available in CMI include: a Zeiss LSM 880 NLO multiphoton/confocal/Airyscan, Olympus Fluoview FV1200 intravital multiphoton, Zeiss LSM 510 confocal, Olympus Fluoview FV 10i confocal, BD BioSciences CARV II spinning disk confocal, and Cytation 5 Cell Imaging Multi-Mode Imager. Microscopes are equipped with environmental chambers for temperature and CO2/O2 control to allow non-destructive high-resolution 3D imaging of living cells and organisms. Applications include: 1) live cell imaging of parameter-sensitive fluorophores to monitor ions, electrical potentials, radical generation, cell viability (apoptosis and necrosis), and fluorescent protein labeling; 2) imaging of tissue sections for immunocytochemistry and fluorescent protein distribution; 3) fluorescence resonance energy transfer (FRET) and Duolink to quantify intermolecular interactions; 4) intravital microscopy to monitor microcirculation, leukocyte margination, mitochondrial polarization, and other factors in living animals; and 5) automated microplate microscopy for high-content screening of compound libraries. Imaging workstations run Imaris, Huygens, ImageJ FIJI, and other software for image processing and analysis. Ancillary equipment includes culture hoods, incubators, and fluorescence and absorbance plate readers. CMI services include consultation and hands-on training and assistance to students, postdoctoral fellows, technical personnel, and both junior and senior investigators. CMI also organizes seminars, invites leading microscope manufacturers for instrument demos, and presents a biennial Charleston Workshop on Light Microscopy for The Biosciences that provides a solid introduction to the concepts and practical applications of light microscopy relevant to modern cell and molecular biology, which will next be held June 11-16, 2022. In the current cycle, CMI supported research in 59 HCC member labs, leading to 114 publications. Under the leadership of John J. Lemasters, MD, PhD (DCT), a national leader in mitochondria and energetics biology, CMI has grown to become a superb scientific resource for the visualization and quantification of cellular processes. The future goals of CMI are to continue providing outstanding services to HCC members and to expand CMI’s super-resolution and image analysis and processing capabilities to ensure that HCC members can perform top-level research.