Routine single particle CryoEM sample and grid characterization by tomography

Abstract
Data availability
Article and author information
Metrics

Abstract

Single particle cryo-electron microscopy (cryoEM) is often performed under the assumption that particles are not adsorbed to the air-water interfaces and in thin, vitreous ice. In this study, we performed fiducial-less tomography on over 50 different cryoEM grid/sample preparations to determine the particle distribution within the ice and the overall geometry of the ice in grid holes. Surprisingly, by studying particles in holes in 3D from over 1,000 tomograms, we have determined that the vast majority of particles (approximately 90%) are adsorbed to an air-water interface. The implications of this observation are wide-ranging, with potential ramifications regarding protein denaturation, conformational change, and preferred orientation. We also show that fiducial-less cryo-electron tomography on single particle grids may be used to determine ice thickness, optimal single particle collection areas and strategies, particle heterogeneity, and de novo models for template picking and single particle alignment.

Data availability

Several representative tilt-series from the datasets have been deposited to the Electron Microscopy Data Bank (EMDB) in the form of binned by 4 or 8 tomograms and to the Electron Microscopy Pilot Image Archive (EMPIAR) in the form of unaligned tilt-series images (one including super-resolution frames), Appion-Protomo tilt-series alignment runs, and aligned tilt-series stacks.Protomo estimations for the orientation of the local ice normal based on the tilt-series alignment of the particles in the ice, which includes potential systematic stage and beam axis error, are available in all deposited EMPIAR datasets as a plot located: protomo_alignments/tiltseries####/media/angle_refinement/series####_orientation.gifA Docker-based version of Appion-Protomo fiducial-less tilt-series alignment is available at http://github.com/nysbc/appion-protomo.

The following data sets were generated

1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of hemagglutinin single particle
EMPIAR-10129.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10129
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of rabbit muscle aldolase single particle super-res
EMPIAR-10130.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10130
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of rabbit muscle aldolase single particle
EMPIAR-10131.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10131
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of glutamate dehydrogenase single particle
EMPIAR-10132.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10132
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of glutamate dehydrogenase single particle
EMPIAR-10133.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10133
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of glutamate dehydrogenase + 0.001% DDM single particle
EMPIAR-10134.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10134
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of DNAB helicase-helicase loader single particle
EMPIAR-10135.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10135
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin single particle
EMPIAR-10136.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10136
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin single particle
EMPIAR-10137.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10137
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin single particle
EMPIAR-10138.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10138
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin single particle
EMPIAR-10139.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10139
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin single particle
EMPIAR-10140.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10140
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of apoferritin with 0.5 mM TCEP single particle
EMPIAR-10141.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10141
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Phase plate cryoET of T20S proteasome single particle
EMPIAR-10142.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10142
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of T20S proteasome single particle
EMPIAR-10143.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10143
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of T20S proteasome single particle
EMPIAR-10144.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10144
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) CryoET of Mtb 20S proteasome single particle
EMPIAR-10145.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10145
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Hemagglutinin on a carbon nanowire grid plunged with Spotiton
EMD-7135.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7135
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Rabbit muscle aldolase on a gold nanowire grid plunged with Spotiton
EMD-7138.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7138
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Rabbit muscle aldolase on a carbon nanowire grid plunged with Spotiton
EMD-7139.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7139
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Protein in nanodisc on a gold nanowire grid plunged with Spotiton
EMD-7140.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7140
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Glutamate dehydrogenase on a holey carbon grid
EMD-7141.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7141
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Glutamate dehydrogenase on a holey carbon grid
EMD-7142.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7142
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) GDH + 0.001% DDM on a carbon nanowire grid plunged with Spotiton
EMD-7143.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7143
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) DNAB helicase-helicase loader on a gold Quantifoil grid
EMD-7144.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7144
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin on a gold nanowire grid plunged with Spotiton
EMD-7145.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7145
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin on a gold nanowire grid plunged with Spotiton
EMD-7146.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7146
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin on a holey carbon nanowire grid plunged with Spotiton
EMD-7147.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7147
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin on a holey carbon nanowire grid plunged with Spotiton
EMD-7148.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7148
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin on a holey gold nanowire grid plunged with Spotiton
EMD-7149.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7149
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Apoferritin with 0.5 mM TCEP on a carbon nanowire grid plunged with Spotiton
EMD-7150.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7150
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) T20S proteasome on a holey carbon grid
EMD-7151.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7151
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) T20S proteasome on a holey carbon grid
EMD-7152.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7152
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) T20S proteasome on a gold Quantifoil grid
EMD-7153.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7153
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2017) Mtb 20S proteasome on a carbon nanowire grid plunged with Spotiton
EMD-7154.

https://www.ebi.ac.uk/pdbe/entry/emdb/EMD-7154
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2018) Rabbit muscle aldolase single particle
EMPIAR-10187.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10187
1. Noble AJ
2. Dandey VP
3. Wei H
4. Brasch J
5. Chase J
6. Acharya P
7. Tan YZ
8. Zhang Z
9. Kim LY
10. Scapin G
11. Rapp M
12. Eng ET
13. Rice WJ
14. Cheng A
15. Negro CJ
16. Shapiro L
17. Kwong PD
18. Jeruzalmi D
19. des Georges A
20. Potter CS
21. Carragher B
(2018) T20S proteasome single particle
EMPIAR-10188.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10188

Article and author information

Author details

Alex J Noble

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-8634-2279
Venkata P Dandey

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Hui Wei

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Julia Brasch

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Jillian Chase

Department of Chemistry and Biochemistry, City College of New York, New York, United States

Competing interests
The authors declare that no competing interests exist.
Priyamvada Acharya

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Yong Zi Tan

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-6656-6320
Zhening Zhang

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Laura Y Kim

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Giovanna Scapin

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Micah Rapp

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Edward T Eng

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-8014-7269
William J Rice

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Anchi Cheng

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Carl J Negro

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.
Lawrence Shapiro

Department of Biochemistry and Molecular Biophysics, Columbia University, New York, United States

Competing interests
The authors declare that no competing interests exist.
Peter D Kwong

Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, United States

Competing interests
The authors declare that no competing interests exist.
David Jeruzalmi

Department of Chemistry and Biochemistry, City College of New York, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5886-1370
Amedee des Georges

Department of Chemistry and Biochemistry, City College of New York, New York, United States

Competing interests
The authors declare that no competing interests exist.
Clinton S Potter

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-2394-0831
Bridget Carragher

National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, United States

For correspondence
bcarr@nysbc.org

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-0624-5020

Funding

Simons Foundation (SF349247)

Clinton S Potter
Bridget Carragher

National Institutes of Health (R01 GM084162)

David Jeruzalmi

New York State Foundation for Science, Technology and Innovation

Clinton S Potter
Bridget Carragher

National Institute of General Medical Sciences (GM103310)

Clinton S Potter
Bridget Carragher

Agouron Institute (F00316)

Clinton S Potter
Bridget Carragher

National Institutes of Health (S10 OD019994-01)

Clinton S Potter
Bridget Carragher

National Institute on Minority Health and Health Disparities (5G12MD007603-30)

David Jeruzalmi

National Institute of Allergy and Infectious Diseases (Intramural Funding from the Vaccine Research Center)

Peter D Kwong

Agency for Science, Technology and Research

Yong Zi Tan

National Institutes of Health (R01-MH1148175)

Lawrence Shapiro

The authors declare that the funders played no role in this work, including the experimental design, data collection, or data analysis.

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.