Cryo-electron microscopy (cryo-EM) is a vital structural biology tool that enables the determination of three-dimensional structures of large biological macromolecules, macromolecular complexes, and cell components of biomedical interest, which bridges the gap of complementary structural biology tools such as X -ray crystallography and nuclear magnetic resonance spectroscopy (NMR). The resolution of singl -particle cryo-EM has dramatically improved in the last couple of years and cryo-EM is now an essential technique for structural biology research. When operating in “molecular movie mode” the Titan Krios cryo-EM with Falcon II direct electron detector will produce major data volumes, generating terabytes of images per day. The data produced will be a significant asset to Australian and international researchers and will permit the determination of structures of large complex proteins that resist characterization through approaches such as X-ray crystallography. The data storage made available through this proposal will allow researchers to more easily capture, share and publish images produced using the high-end microscopes of the Clive and Vera Ramaciotti Centre for Structural Cryo-Electron Microscopy. More broadly, this undertaking is important to support Australian investment in characterization, providing the missing fabric to collect, collate, and collaborate over data. It will form the basis for a sophisticated workflow, developed under NeCTAR’s Characterisation Virtual Laboratory, that includes significant Australian research infrastructure like the NeCTAR Cloud, MASSIVE, RDSI and VicNode. This Collection will make imaging data from research grants totalling over $100M and instrument grants totalling more than $7M accessible to researchers and provide a mechanism to make the primary research data underlying published research broadly available. This includes data produced by the TitanTM Krios cryo TEM at the Clive and Vera Ramaciotti Centre for Structural Cryo-Electron Microscopy (Monash, in collaboration with WEHI, La Trobe University, the University of Melbourne, Burnet Institute and the Peter MacCallum Cancer Centre). This collection will underpin the $27.9M ARC CoE in Advanced Molecular Imaging, which includes collaborators from Monash University, UNSW, UQ, The University of Melbourne, La Trobe University, Australian Synchrotron, University of Warwick, ANSTO, Deutsches ElektronenJSynchrotron, Leica Microsystems Pty Ltd and Carl Zeiss Pty Ltd.