S'identifier

Simons Electron Microscopy Center, New York Structural Biology Center

3 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.

Biology

Leveraging Virtual Reality for Immersive Segmentation and Analysis of Cryo-Electron Tomography Data

Carissa Chestnut^*1,2, Jake D. Johnston^*2,3, Marcus Velazquez^*1,2, Kasahun Neselu², Edward T. Eng²

¹Stony Brook University, ²Simons Electron Microscopy Center, New York Structural Biology Center, ³Department of Physiology and Cellular Biophysics, Columbia University

Cryo-electron tomography (cryo-ET) enables 3D visualization of cellular ultrastructure at nanometer resolution, but manual segmentation remains time-consuming and complex. We present a novel workflow that integrates advanced virtual reality software for segmenting cryo-ET tomograms, showcasing its effectiveness through the segmentation of mitochondria in mammalian cells.