MicroED with the Falcon III direct electron detector
Microcrystal electron diffraction (MicroED) combines crystallography and electron cryo-microscopy (cryo-EM) into a method that is applicable to high-resolution structure determination. In MicroED, nanosized crystals, which are often intractable using other techniques, are probed by high-energy elect...
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International Union of Crystallography
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doaj-63f09f2e6aeb443eb21cc6704feb04912020-11-25T02:19:02ZengInternational Union of CrystallographyIUCrJ2052-25252019-09-016592192610.1107/S2052252519010583fq5007MicroED with the Falcon III direct electron detectorJohan Hattne0Michael W. Martynowycz1Pawel A. Penczek2Tamir Gonen3Howard Hughes Medical Institute, Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USAHoward Hughes Medical Institute, Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USADepartment of Biochemistry and Molecular Biology, The University of Texas McGovern Medical School, Houston, TX 77030, USAHoward Hughes Medical Institute, Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USAMicrocrystal electron diffraction (MicroED) combines crystallography and electron cryo-microscopy (cryo-EM) into a method that is applicable to high-resolution structure determination. In MicroED, nanosized crystals, which are often intractable using other techniques, are probed by high-energy electrons in a transmission electron microscope. Diffraction data are recorded by a camera in movie mode: the nanocrystal is continuously rotated in the beam, thus creating a sequence of frames that constitute a movie with respect to the rotation angle. Until now, diffraction-optimized cameras have mostly been used for MicroED. Here, the use of a direct electron detector that was designed for imaging is reported. It is demonstrated that data can be collected more rapidly using the Falcon III for MicroED and with markedly lower exposure than has previously been reported. The Falcon III was operated at 40 frames per second and complete data sets reaching atomic resolution were recorded in minutes. The resulting density maps to 2.1 Å resolution of the serine protease proteinase K showed no visible signs of radiation damage. It is thus demonstrated that dedicated diffraction-optimized detectors are not required for MicroED, as shown by the fact that the very same cameras that are used for imaging applications in electron microscopy, such as single-particle cryo-EM, can also be used effectively for diffraction measurements.http://scripts.iucr.org/cgi-bin/paper?S2052252519010583microcrystal electron diffractionMicroEDFalcon IIIdirect electron detectors |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Johan Hattne Michael W. Martynowycz Pawel A. Penczek Tamir Gonen |
spellingShingle |
Johan Hattne Michael W. Martynowycz Pawel A. Penczek Tamir Gonen MicroED with the Falcon III direct electron detector IUCrJ microcrystal electron diffraction MicroED Falcon III direct electron detectors |
author_facet |
Johan Hattne Michael W. Martynowycz Pawel A. Penczek Tamir Gonen |
author_sort |
Johan Hattne |
title |
MicroED with the Falcon III direct electron detector |
title_short |
MicroED with the Falcon III direct electron detector |
title_full |
MicroED with the Falcon III direct electron detector |
title_fullStr |
MicroED with the Falcon III direct electron detector |
title_full_unstemmed |
MicroED with the Falcon III direct electron detector |
title_sort |
microed with the falcon iii direct electron detector |
publisher |
International Union of Crystallography |
series |
IUCrJ |
issn |
2052-2525 |
publishDate |
2019-09-01 |
description |
Microcrystal electron diffraction (MicroED) combines crystallography and electron cryo-microscopy (cryo-EM) into a method that is applicable to high-resolution structure determination. In MicroED, nanosized crystals, which are often intractable using other techniques, are probed by high-energy electrons in a transmission electron microscope. Diffraction data are recorded by a camera in movie mode: the nanocrystal is continuously rotated in the beam, thus creating a sequence of frames that constitute a movie with respect to the rotation angle. Until now, diffraction-optimized cameras have mostly been used for MicroED. Here, the use of a direct electron detector that was designed for imaging is reported. It is demonstrated that data can be collected more rapidly using the Falcon III for MicroED and with markedly lower exposure than has previously been reported. The Falcon III was operated at 40 frames per second and complete data sets reaching atomic resolution were recorded in minutes. The resulting density maps to 2.1 Å resolution of the serine protease proteinase K showed no visible signs of radiation damage. It is thus demonstrated that dedicated diffraction-optimized detectors are not required for MicroED, as shown by the fact that the very same cameras that are used for imaging applications in electron microscopy, such as single-particle cryo-EM, can also be used effectively for diffraction measurements. |
topic |
microcrystal electron diffraction MicroED Falcon III direct electron detectors |
url |
http://scripts.iucr.org/cgi-bin/paper?S2052252519010583 |
work_keys_str_mv |
AT johanhattne microedwiththefalconiiidirectelectrondetector AT michaelwmartynowycz microedwiththefalconiiidirectelectrondetector AT pawelapenczek microedwiththefalconiiidirectelectrondetector AT tamirgonen microedwiththefalconiiidirectelectrondetector |
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