The Collection of High-Resolution Electron Diffraction Data
A number of atomic-resolution structures of membrane proteins (better than 3Å resolution) have been determined recently by electron crystallography. While this technique was established more than 40 years ago, it is still in its infancy with regard to the two-dimensional (2D) crystallization, data collection, data analysis, and protein structure determination. In terms of data collection, electron crystallography encompasses both image acquisition and electron diffraction data collection. Other chapters in this volume outline protocols for image collection and analysis. This chapter, however, outlines detailed protocols for data collection by electron diffraction. These include microscope setup, electron diffraction data collection, and troubleshooting.
Key wordsElectron cryomicroscopy (Cryo-EM) Electron crystallography Electron diffraction
We thank Noah Shuart (University of Washington) for critically reading this manuscript. Research in the Gonen laboratory is supported by the American Diabetes Association Award # 1-09-CD-05 and by the National Institutes of Health R01GM079233 and U54GM094598 as well as the Howard Hughes Medical Institute.