2017Moriya Sphire

From 3DEM-Methods
Jump to: navigation, search

Citation

Moriya, T.; Saur, M.; Stabrin, M.; Merino, F.; Voicu, H.; Huang, Z.; Penczek, P. A.; Raunser, S. & Gatsogiannis, C. High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE. Journal of visualized experiments : JoVE, 2017

Abstract

SPHIRE (SPARX for High-Resolution Electron Microscopy) is a novel open-source, user-friendly software suite for the semi-automated processing of single particle electron cryo-microscopy (cryo-EM) data. The protocol presented here describes in detail how to obtain a near-atomic resolution structure starting from cryo-EM micrograph movies by guiding users through all steps of the single particle structure determination pipeline. These steps are controlled from the new SPHIRE graphical user interface and require minimum user intervention. Using this protocol, a 3.5 Å structure of TcdA1, a Tc toxin complex from Photorhabdus luminescens, was derived from only 9500 single particles. This streamlined approach will help novice users without extensive processing experience and a priori structural information, to obtain noise-free and unbiased atomic models of their purified macromolecular complexes in their native state.

Keywords

Links

https://www.jove.com/video/55448/high-resolution-single-particle-analysis-from-electron-cryo

Related software

Related methods

Comments