Microcrystal Electron Diffraction of Small Molecules

A detailed protocol for preparing small molecule samples for microcrystal electron diffraction (MicroED) experiments is described. MicroED has been developed to solve structures of proteins and small molecules using standard electron cryo-microscopy (cryo-EM) equipment. In this way, small molecules, peptides, soluble proteins, and membrane proteins have recently been determined to high resolutions. Protocols are presented here for preparing grids of small-molecule pharmaceuticals using the drug carbamazepine as an example. Protocols for screening and collecting data are presented. Additional steps in the overall process, such as data integration, structure determination, and refinement are presented elsewhere. The time required to prepare the small-molecule grids is estimated to be less than 30 min.

Automated summary

The detailed protocol for preparing small molecule samples for microcrystal electron diffraction (MicroED) experiments is described in this article, which has been developed to solve structures of proteins and small molecules using standard electron cryo-microscopy equipment. Recent advancements have shown that this method can be applied to a variety of small molecules and protein structures.