Advances of 3D microcrystals electron diffraction for transmembrane protein structure determination

Electron diffraction of 3D microcrystals (3D ED/MicroED) is an emerging technique that uses electrons to determine proteins structure from their nano-sized 3D crystals, thus overcoming the limitations on crystal-size dimensions generally imposed by x-ray or neutron diffraction. The strong interactions of electrons with matter have the potential to reveal the location of light atoms (e.g., hydrogens), revealing interaction networks, and to provide information about the charge states of the atoms. Recent advances in MicroED sample preparation and data collection strategies have led to structural determination of several known and unknown protein structures, including challenging targets such as membrane proteins. In this review, we provide an overview of the recent advances of MicroED including sample preparation and data collection.