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Bridget Carragher (Chair), Jose-Maria Carazo, Wen Jiang, John Rubinstein, Peter Rosenthal, Fei Sun, Janet Vonck, Wah Chiu, Cathy Lawson, Ardan Patwardhan
All members of the Scientific Community--at all levels of experience--are invited to participate as Challengers, and/or as Assessors.
In the Challenge Phase (July 2015-Apr 2016), 27 participants created and submitted 66 reconstructions of the challenge targets using the supplied raw image data. Challengers were encouraged to perform their own movie frame alignment, frame summation, and particle picking. Alternately, they could begin with pre-aligned, summed images and/or original author-provided particle positions. For each submission, challengers filled out a questionnaire (overview) and provided the following data:
In the Blinded Assessment Phase (Nov 2016-Apr 2017), six groups have contributed reports. Following initial review period by the map committee, the challenge data and files are now publicly available (entry authorship and software suppressed) for anyone to assess. Assessment methods could include statistical analyses, resolution estimation, or coordinate model fitting. A few suggestions gathered from software developers are summarized below. The intention is to enable comparisons of the various packages available and their options in a positive spirit. During this period, assessors are strongly encouraged to share their plans and short result summaries on this website using the Assessment Registration Form. Assessment results will be more fully presented and discussed via a workshop (early October 2017) as well as via manuscript submissions to a Journal special issue.
We've gathered some suggestions here about how to proceed with comparisons of the map submissions.* These are not meant to be prescriptive; results from other approaches are also welcome.
FSC curves based on provided half-maps and masks have been prepared for each map challenge entry (link). In most cases the results are consistent with or very close to the submitter-reported resolution, but this initial analysis cannot be used to directly compare submissions, because of differences in masking and map sizes, and thus convolution effects. FSC is a fundamental similarity metric, but its use in standard cryoEM practice has been problematic because of the maps being compared. Many suggestions were made on how to carry out follow-up analyses:
Images of each map both by itself and aligned to a common model are provided for reference (link), but further investigation is warranted, as variations in density appearance may be due to differences in power spectra and/or filtering/sharpening schemes. Some suggestions:
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*suggestion credits: Maya Holmdahl, Roberto Marabini, Sjors Scheres, Bernard Heymann, Niko Grigorieff, Pawel Penczek, Ed Egelman, Steve Ludtke, Scott Stagg, Marin van Heel
JAN-JUN 2015 |
DEVELOPMENT PHASE |
Jan-May | Map Committee meets monthly to identify challenge targets, goals, and parameters |
Mar-May | Requests to 3DEM community members for public deposition of raw image Datasets; website development |
July 1 | Raw image data for all targets available for download at EMPIAR |
JUL 2015-APR 2016 |
CHALLENGE PHASE |
July | Pre-Challenge Announcement, Challenger and Assessor Registration Opens |
August 1 | Map Submission Opens |
Nov 9-20 | Registered Participants may apply for SDSC Gordon Supercomputer Usage |
April 1-2 | Map Challenge Committee satellite discussion @ International CryoEM Image Analysis Symposium (Lake Tahoe, CA) |
Apr 15 | Map Submission Closes @ 21:00 UTC |
2016-2017 | ASSESSMENT PHASE |
May-August | Challenge Data initial assessments, metadata extraction, preparation for release (Map Committee) |
Sept-Oct |
Review Period (Map Committee) |
4 Nov 2016-30 Apr 2017 | Assessors invited to perform analyses and comment on Released Data (Blinded) |
6 June 2017 | Map Submission Data UnBlinded |
June - Sept 2017 | Analysis of Assessments with full metadata |
Oct 6-8 2017 |
2 day Workshop for all challenge participants -- Committee, Challengers, Assessors |
Post-workshop | Challenge Writeups (multiple articles) for Journal Special Issue |
Six challenge targets are based on recently described 3DEM single particle structure determinations with data collected as multiple-frames-per-second movies, using the latest generation of detectors. One additional target is based on simulated (in silico) images. For each experimental target, the original raw micrograph movie frames are data available for download at EMPIAR, PDBe's raw 3DEM image data archive. Summed image data are also available, either as full micrographs or as picked particle stacks. In one case aligned frames are also deposited. Particle positions and defocus values from the raw data depositors are also available for download and may optionally be used by challengers in their reconstructions.
target | 1. GroEL in silico | 2. T20S Proteasome | 3. Apo-Ferritin | 4. TRPV1 Channel | 5. 80S Ribosome | 6. Brome Mosaic Virus | 7. β-Galactosidase |
Reference EMDB map entry |
-- |
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Primary Citation | Vulovic et al | Campbell et al | Russo & Passmore | Liao et al | Wong et al | Wang et al | Bartesaghi et al |
Reported Resolution (Å) | ~3 | 2.8 | 4.7 | 3.3 | 3.2 | 3.8 | 3.2 |
Reference Model PDB entry |
4hel |
1yar |
4v1w |
3j5p |
3j79/3j7a |
3j7l |
5a1a |
Benchmark Storage Size |
2 GB |
2000 GB |
181 GB |
6300 GB |
2000 GB |
460 GB |
550 GB |
EMPIAR ID(s) data can also be downloaded from Chinese Academy of Sciences |
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Raw Frames |
n.a. | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ |
Aligned Frames | ✔ | ||||||
Summed Micrographs |
✔ | ✔ | ✔ | ✔ | |||
Summed Particle Stacks | ✔ | ✔ | ✔ | ✔ | |||
Initial Particle Coordinates (directory link) |
spider FAQ | eman-box | eman-box | ||||
Final Particle Coordinates (direct file link) |
relion-star | relion-star | relion-star | ||||
Particle coordinates in EMX format; python script used for conversion files contributed by Roberto Marabini and Jose Maria Carazo |
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Imposed Symmetry |
Dihedral (D7) None (C1) |
Dihedral (D7) | Octahedral (O) | Cyclic (C4) | None (C1) | Icosahedral (I) | Dihedral (D2) |
Sample MW (MDa) | 0.8 | 0.7 | 0.44 | 0.3 | 4.2 | 4.6 | 0.47 |
Unique MW (kDa) | 57 | 50 | 20 | 80 | 4200 | 80 | 120 |
Microscope | -- | Titan Krios | Polara 300 | Polara 300 | Polara 300 | JEOL3200FSC | Titan Krios |
Voltage(kV) | 300 | 300 | 300 | 300 | 300 | 300 | 300 |
Cs (mm) |
2.7 | 2.7 | 2.7 FAQ | 2.0 | 2.0 | 4.1 | 2.7 |
Detector | Falcon I | K2 | Falcon II | K2 | Falcon II | DE12 | K2 |
Frame Sampling (Å/pixel) | 1.42 | 0.6575 | 1.346 | 1.22 FAQ | 1.34 | 0.99 | 0.64 |
total dose (e-/Å2) | 50 | 53 | 16 | 41 | 20 | 52 | 45 |
dose per frame (e-/Å2) | -- | 1.4 | 0.95 | 1.37 | 1 | 1.4 | 1.2 |
frame rate (f/s) | -- | 5 | 17 | 5 | 16 | 25 | 2.5 |
frame alignment method | -- | UCSF | not performed | UCSF | Statistical | DE script | cross-correlation script |
Particle selection method | -- | Appion-FindEM | EMAN2 | SamViewer | EMAN2 | EMAN2 | Gaussian correlation |
Number of Particles | 10000 | 49954 | 483 | 35645 | 105247 | 30000 | 11726 |
Particle/Map Sampling (Å/pixel) | 1.42 | 0.98 | 1.346 | 1.22 | 1.34 | 0.99 | 0.64 |
Raw Data Contributors (Thank You!) | Yuchen Deng, Fei Sun | Melody Campbell, Bridget Carragher | Chris Russo, Lori Passmore | Jean-Paul Armache, Maofu Liao, Yifan Cheng | Xiaochen Bai, Sjors Scheres | Zhao Wang, Wah Chiu | Alberto Bartesaghi, Sriram Subramaniam |
EMDataResource Validation Challenges are supported by NIH National Institute of General Medical Sciences
Please send your challenge questions, comments and feedback to challenges@emdataresource.org
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