MicroED

Cryo-electron microscopy (cryo-EM) methods promise new life to high-throughput macromolecular structure determination. Cryo-EM overcomes the fundamental barrier to X-ray diffraction determination of macromolecular complexes: growing X-ray grade crystals. Fortunately, the emergence of microcrystal electron diffraction (MicroED) facilitates the determination of new protein structures at atomic resolution from vanishingly small crystals.

MicroED exploits the strong interaction between electrons and nano-scale three-dimensional crystals and takes advantage of emerging cryo-EM instrumentation coupled to established crystallographic methods.

Gonen pointing at computer in lab
Tamir Gonen on MicroED: In this interview, Tamir Gonen (UCLA) outlines the history of MicroED and its relationship to other methods for structure determination. While originally motivated by the desire to study membrane proteins, the up-and-coming method has now been applied to many different samples that often are not amenable to structural analysis by other means.

Watch the Video: Interview with Dr. Tamir Gonen

MicroED exploits the strong interaction between electrons and nano-scale three-dimensional crystals and takes advantage of emerging cryo-EM instrumentation coupled to established crystallographic methods.

MicroED on a globular protein
MicroED on a globular protein: proteinase K, a 30 kDa hydrolase which crystallizes in a tetragonal spacegroup, has been used extensively for methods development on MicroED.

Watch the Video: MicroED on a globular protein

Lab technician loading sample into TEM

Watch video: Microcrystal Electron Diffraction of Small Molecules

Suggested Further Reading

2021

Martynowycz, Michael W; Clabbers, Max T B; Unge, Johan; Hattne, Johan; Gonen, Tamir
Benchmarking the ideal sample thickness in cryo-EM Journal Article
In: Proc Natl Acad Sci U S A, vol. 118, no. 49, 2021, ISSN: 1091-6490.

2020

Zhu, Lan; Bu, Guanhong; Jing, Liang; Shi, Dan; Lee, Ming-Yue; Gonen, Tamir; Liu, Wei; Nannenga, Brent L
Structure Determination from Lipidic Cubic Phase Embedded Microcrystals by MicroED Journal Article
In: Structure, vol. 28, no. 10, pp. 1149–1159.e4, 2020, ISSN: 1878-4186.

2018

Jones, Christopher G; Martynowycz, Michael W; Hattne, Johan; Fulton, Tyler J; Stoltz, Brian M; Rodriguez, Jose A; Nelson, Hosea M; Gonen, Tamir
The CryoEM Method MicroED as a Powerful Tool for Small Molecule Structure Determination Journal Article
In: ACS Cent Sci, vol. 4, no. 11, pp. 1587–1592, 2018, ISSN: 2374-7943.

2013

Shi, Dan; Nannenga, Brent L; Iadanza, Matthew G; Gonen, Tamir
Three-dimensional electron crystallography of protein microcrystals Journal Article
In: Elife, vol. 2, pp. e01345, 2013, ISSN: 2050-084X.

Suggested Reviews

2021

Danelius, Emma; Gonen, Tamir
Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED Journal Article
In: Methods Mol Biol, vol. 2305, pp. 323–342, 2021, ISSN: 1940-6029.

Martynowycz, Michael W; Gonen, Tamir
Studying Membrane Protein Structures by MicroED Journal Article
In: Methods Mol Biol, vol. 2302, pp. 137–151, 2021, ISSN: 1940-6029.

Mu, Xuelang; Gillman, Cody; Nguyen, Chi; Gonen, Tamir
An Overview of Microcrystal Electron Diffraction (MicroED) Journal Article
In: Annu Rev Biochem, vol. 90, pp. 431–450, 2021, ISSN: 1545-4509.

2017

Liu, Shian; Hattne, Johan; Reyes, Francis E; Sanchez-Martinez, Silvia; de la Cruz, M Jason; Shi, Dan; Gonen, Tamir
Atomic resolution structure determination by the cryo-EM method MicroED Journal Article
In: Protein Sci, vol. 26, no. 1, pp. 8–15, 2017, ISSN: 1469-896X.

2016

Nannenga, Brent L; Gonen, Tamir
MicroED opens a new era for biological structure determination Journal Article
In: Curr Opin Struct Biol, vol. 40, pp. 128–135, 2016, ISSN: 1879-033X.
 

2014

Nannenga, Brent L; Shi, Dan; Leslie, Andrew G W; Gonen, Tamir
High-resolution structure determination by continuous-rotation data collection in MicroED Journal Article
In: Nat Methods, vol. 11, no. 9, pp. 927–930, 2014, ISSN: 1548-7105.

Protocols

2021

Martynowycz, Michael W; Gonen, Tamir
Protocol for the use of focused ion-beam milling to prepare crystalline lamellae for microcrystal electron diffraction (MicroED) Journal Article
In: STAR Protoc, vol. 2, no. 3, pp. 100686, 2021, ISSN: 2666-1667.

2017

de la Cruz, M Jason; Hattne, Johan; Shi, Dan; Seidler, Paul; Rodriguez, Jose; Reyes, Francis E; Sawaya, Michael R; Cascio, Duilio; Weiss, Simon C; Kim, Sun Kyung; Hinck, Cynthia S; Hinck, Andrew P; Calero, Guillermo; Eisenberg, David; Gonen, Tamir
Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED Journal Article
In: Nat Methods, vol. 14, no. 4, pp. 399–402, 2017, ISSN: 1548-7105.

2016

Shi, Dan; Nannenga, Brent L; de la Cruz, M Jason; Liu, Jinyang; Sawtelle, Steven; Calero, Guillermo; Reyes, Francis E; Hattne, Johan; Gonen, Tamir
The collection of MicroED data for macromolecular crystallography Journal Article
In: Nat Protoc, vol. 11, no. 5, pp. 895–904, 2016, ISSN: 1750-2799.