Simons Electron Microscopy Center, New York Structural Biology Center 2 articles published in JoVE Biochemistry Cryo-Electron Microscopy Screening Automation Across Multiple Grids Using Smart Leginon Anjelique Sawh-Gopal1,2, Aygul Ishemgulova*1, Eugene Y. D. Chua*1, Mahira F. Aragon*1, Joshua H. Mendez1, Edward T. Eng1, Alex J. Noble1 1Simons Electron Microscopy Center, New York Structural Biology Center, 2Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology Cryo-electron microscopy (cryoEM) multi-grid screening is often a tedious process that demands hours of attention. This protocol shows how to set up a standard Leginon collection and Smart Leginon Autoscreen to automate this process. This protocol can be applied to the majority of cryoEM holey foil grids. Biology Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton William C. Budell1, Luis Allegri2, Venkat Dandey1, Clinton S. Potter1, Bridget Carragher1 1The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, 2piTree Software The protocol presented here describes the use of Spotiton, a novel robotic system, to deliver two samples of interest onto a self-wicking, nanowire grid that mix for a minimum of 90 ms prior to vitrification in liquid cryogen.
