The University of Texas Medical School at Houston 1 article published in JoVE Biochemistry High-resolution Single Particle Analysis from Electron Cryo-microscopy Images Using SPHIRE Toshio Moriya1, Michael Saur1, Markus Stabrin1, Felipe Merino1, Horatiu Voicu2, Zhong Huang2, Pawel A. Penczek2, Stefan Raunser1, Christos Gatsogiannis1 1Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, 2Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston This paper presents a protocol for processing cryo-EM images using the software suite SPHIRE. The present protocol can be applied for nearly all single particle EM projects that target near-atomic resolution.