Influence of electron dose rate on electron counting images recorded with the K2 camera. 2013

Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
The Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, CA 94158, United States.

A recent technological breakthrough in electron cryomicroscopy (cryoEM) is the development of direct electron detection cameras for data acquisition. By bypassing the traditional phosphor scintillator and fiber optic coupling, these cameras have greatly enhanced sensitivity and detective quantum efficiency (DQE). Of the three currently available commercial cameras, the Gatan K2 Summit was designed specifically for counting individual electron events. Counting further enhances the DQE, allows for practical doubling of detector resolution and eliminates noise arising from the variable deposition of energy by each primary electron. While counting has many advantages, undercounting of electrons happens when more than one electron strikes the same area of the detector within the analog readout period (coincidence loss), which influences image quality. In this work, we characterized the K2 Summit in electron counting mode, and studied the relationship of dose rate and coincidence loss and its influence on the quality of counted images. We found that coincidence loss reduces low frequency amplitudes but has no significant influence on the signal-to-noise ratio of the recorded image. It also has little influence on high frequency signals. Images of frozen hydrated archaeal 20S proteasome (~700 kDa, D7 symmetry) recorded at the optimal dose rate retained both high-resolution signal and low-resolution contrast and enabled calculating a 3.6 Å three-dimensional reconstruction from only 10,000 particles.

UI MeSH Term Description Entries
D008958 Models, Molecular Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures. Molecular Models,Model, Molecular,Molecular Model
D010450 Endopeptidases A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS. Endopeptidase,Peptide Peptidohydrolases
D000465 Algorithms A procedure consisting of a sequence of algebraic formulas and/or logical steps to calculate or determine a given task. Algorithm
D013822 Thermoplasma A genus of facultatively anaerobic heterotrophic archaea, in the order THERMOPLASMALES, isolated from self-heating coal refuse piles and acid hot springs. They are thermophilic and can grow both with and without sulfur.
D057230 Limit of Detection Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure. Limits of Detection,Detection Limit,Detection Limits
D059629 Signal-To-Noise Ratio The comparison of the quantity of meaningful data to the irrelevant or incorrect data. Ratio, Signal-To-Noise,Ratios, Signal-To-Noise,Signal To Noise Ratio,Signal-To-Noise Ratios
D019843 Archaeal Proteins Proteins found in any species of archaeon. Archaeal Gene Products,Archaeal Gene Proteins,Archaeal Peptides,Gene Products, Archaeal,Gene Proteins, Archaeal
D020285 Cryoelectron Microscopy Electron microscopy involving rapid freezing of the samples. The imaging of frozen-hydrated molecules and organelles permits the best possible resolution closest to the living state, free of chemical fixatives or stains. Electron Cryomicroscopy,Cryo-electron Microscopy,Cryo electron Microscopy,Cryo-electron Microscopies,Cryoelectron Microscopies,Cryomicroscopies, Electron,Cryomicroscopy, Electron,Electron Cryomicroscopies,Microscopies, Cryo-electron,Microscopies, Cryoelectron,Microscopy, Cryo-electron,Microscopy, Cryoelectron
D020836 Protein Structure, Quaternary The characteristic 3-dimensional shape and arrangement of multimeric proteins (aggregates of more than one polypeptide chain). Quaternary Protein Structure,Protein Structures, Quaternary,Quaternary Protein Structures

Related Publications

Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
November 2015, Journal of structural biology,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
December 2022, Journal of structural biology,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
July 1975, Journal of nuclear medicine : official publication, Society of Nuclear Medicine,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
June 1974, Journal of nuclear medicine : official publication, Society of Nuclear Medicine,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
May 2017, Ultramicroscopy,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
January 1995, IEEE transactions on image processing : a publication of the IEEE Signal Processing Society,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
May 2024, Animals : an open access journal from MDPI,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
June 2018, Proceedings of the National Academy of Sciences of the United States of America,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
January 1994, Journal of nuclear medicine : official publication, Society of Nuclear Medicine,
Xueming Li, and Shawn Q Zheng, and Kiyoshi Egami, and David A Agard, and Yifan Cheng
June 2022, Microscopy (Oxford, England),
Copied contents to your clipboard!