Automation of Continuous-Rotation Data Collection for MicroED
- 2 Citations
- 1.5k Downloads
Abstract
Automated coordination of microscope and camera functions for MicroED data collection simplifies the procedure for robust dataset acquisition and enables unattended sequential collection of many crystal targets. This chapter discusses the prerequisites for an algorithm of data collection automation for continuous-rotation MicroED and presents a practical protocol for achieving this goal using the popular TEM control software program SerialEM.
Key words
MicroED CryoEM Electron microscopy Microcrystal Electron diffraction 3D Electron crystallographyNotes
Acknowledgments
I would like to thank Tamir Gonen for access to a Thermo Fisher Talos Arctica TEM for testing and data collection, and Shian Liu and M. Rhyan Puno for their critical reading of this manuscript. This work is supported by the NIH/NCI Cancer Center Support Grant P30 CA008748 and the Sloan Kettering Institute.
References
- 1.Tan YZ, Cheng A, Potter CS et al (2016) Automated data collection in single particle electron microscopy. Microscopy (Oxf) 65:43–56CrossRefGoogle Scholar
- 2.Mastronarde DN (2005) Automated electron microscope tomography using robust prediction of specimen movements. J Struct Biol 152:36–51CrossRefGoogle Scholar
- 3.Yonekura K, Ishikawa T, Maki-Yonekura S (2019) A new cryo-EM system for electron 3D crystallography by eEFD. J Struct Biol 206:243–253CrossRefGoogle Scholar
- 4.Suloway C, Pulokas J, Fellmann D et al (2005) Automated molecular microscopy: the new Leginon system. J Struct Biol 151:41–60CrossRefGoogle Scholar
- 5.de la Cruz MJ, Martynowycz MW, Hattne J et al (2019) MicroED data collection with SerialEM. Ultramicroscopy 201:77–80CrossRefGoogle Scholar
- 6.Cheng A (2013) Automation of data acquisition in electron crystallography. In: Schmidt-Krey I, Cheng Y (eds) Electron crystallography of soluble and membrane proteins, Methods in Molecular Biology, vol 355. Humana Press, Totowa NJ, pp 307–312CrossRefGoogle Scholar
- 7.Nakamura N, Shimizu Y, Shinkawa T et al (2010) Automated specimen search in cryo-TEM observation with DIFF-defocus imaging. J Electron Microsc 59:299–310CrossRefGoogle Scholar
- 8.Shi D, Nannenga BL, de la Cruz MJ et al (2016) The collection of MicroED data for macromolecular crystallography. Nat Protoc 11:895–904CrossRefGoogle Scholar
- 9.Hattne J, Reyes FE, Nannenga BL et al (2015) MicroED data collection and processing. Acta Crystallogr Sect A Found Adv 71:353–360CrossRefGoogle Scholar