Core B - Advance Imaging Core

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Summary The Proteostasis Advanced Imaging Core (Core B) referred in the Program Project (PP) text as “Image Core” will serve the four projects and the Innovation component of Core D of this PP. The long-term goal is to provide support on image-based studies to understand the cellular and subcellular changes in autophagy dynamics during aging and in the context of Alzheimer disease (AD)-related proteotoxicity. The specific aims of the core are: 1) to provide an imaging resource customized for the study of proteostasis in aging that will assist the PP investigators in the experimental preparation related to: basic microscopy techniques (wide-field microscopy, confocal microscopy, live cell imaging - at Mount Sinai/Einstein) and electron microscopy (at Einstein); 2) to provide a state-of-the art in vivo imaging platform based on two-photon microscopy (Innovation Unit - Mount Sinai) for dynamic analysis of changes in autophagy in brain and peripheral tissues in physiological aging and in disease; 3) to implement clearing imaging techniques and light-sheet microscopy for volume imaging to study autophagy in whole organs (Innovation Unit); 4) to develop STED superresolution imaging methods to study the dynamics of autophagy at the organelle level by nanoscale imaging (Innovation Unit). Components: 1) The Imaging unit provides advice on sample preparation/processing, techniques and procedures for imaging in vivo and fixed samples. Performs electron microscopy (at Einstein) and specialized imaging procedures (at Mount Sinai) such as intravital microscopy, light-sheet imaging, STED superresolution and assists with imaging data interpretation; 2) The innovation unit (at Mount Sinai) will develop and implement procedures specific for analysis of autophagy at the tissue level (light-sheet imaging, intravital microscopy) and at the single molecule level (superresolution imaging) in aging and AD. Services: The core will continue offering i) assistance with regular wide-field microscopy, real-time live cell imaging, confocal and electron microscopy, ii) distributing common reagents (antibodies, probes and fluorescence dyes), iii) sharing detailed protocols for image-based procedures and iv) compiling the image- based information for the PP data base. In addition, during this period the core has now incorporated: i) high- resolution imaging procedures such as intravital two-photon microscopy, superresolution STED imaging and light-sheet microscopy for volume imaging, ii) assistance with sample preparation for these procedures (in vivo mouse preparation, clearing methods) and iii) access to new image analysis software (IMARIS, ARIVIS, Huygens) available at the Microscopy CoRE at Mount Sinai. Relevance: The new technology incorporated in the core now allows dynamic and molecular resolution imaging to study aging and AD-related changes in autophagy at the subcellular level. This information is essential for future implementation of therapeutic approaches aiming at reverting these changes. The centralization of these new procedures optimizes cost efficiency and guarantees standardization and integration of the different groups.