Simons Electron Microscopy Center, New York Structural Biology Center

2 ARTICLES PUBLISHED IN JoVE

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.

Biochemistry

Cryo-Electron Microscopy Screening Automation Across Multiple Grids Using Smart Leginon

Anjelique Sawh-Gopal^1,2, 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.