Journal: J Struct Biol / Year: 2015 Title: Semi-automated selection of cryo-EM particles in RELION-1.3. Authors: Sjors H W Scheres / Abstract: The selection of particles suitable for high-resolution cryo-EM structure determination from noisy micrographs may represent a tedious and time-consuming step. Here, a semi-automated particle ...The selection of particles suitable for high-resolution cryo-EM structure determination from noisy micrographs may represent a tedious and time-consuming step. Here, a semi-automated particle selection procedure is presented that has been implemented within the open-source software RELION. At the heart of the procedure lies a fully CTF-corrected template-based picking algorithm, which is supplemented by a fast sorting algorithm and reference-free 2D class averaging to remove false positives. With only limited user-interaction, the proposed procedure yields results that are comparable to manual particle selection. Together with an improved graphical user interface, these developments further contribute to turning RELION from a stand-alone refinement program into a convenient image processing pipeline for the entire single-particle approach.
History
Deposition
Nov 19, 2014
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Header (metadata) release
Dec 10, 2014
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Map release
Dec 10, 2014
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Update
Mar 11, 2015
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Current status
Mar 11, 2015
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
EMPIAR-10017 (Title: Beta-galactosidase Falcon-II micrographs plus manually selected coordinates by Richard Henderson Data size: 5.3 Data #1: Beta-galactosidase micrographs + coordinates [micrographs - single frame])
Category: CCD / Film or detector model: FEI FALCON II (4k x 4k) / Number real images: 84 / Average electron dose: 24 e/Å2 Details: Every image is the average of 24 raw (unaligned) movie frames. The complete set of electron micrographs used to obtain the density map presented here is available through the Electron ...Details: Every image is the average of 24 raw (unaligned) movie frames. The complete set of electron micrographs used to obtain the density map presented here is available through the Electron Microscopy Pilot Image ARchive (EMPIAR).
Tilt angle min
0
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Model: Tecnai Polara / Image courtesy: FEI Company
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Image processing
Details
The particles were selected using the new semi-automatic particle selection procedures in RELION-1.3. This reconstruction was done to test these procedures.
CTF correction
Details: RELION
Final reconstruction
Applied symmetry - Point group: D2 (2x2 fold dihedral) / Resolution.type: BY AUTHOR / Resolution: 4.2 Å / Resolution method: OTHER / Software - Name: RELION, CTFFIND3 / Number images used: 42755
Final angle assignment
Details: RELION
FSC plot (resolution estimation)
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