References
1. Nannenga BL (2020) MicroED methodology
and development. Struct Dyn 7(1):014304.
https://doi.org/10.1063/1.5128226
2. Nannenga BL, Gonen T (2019) The cryo-EM
method microcrystal electron diffraction
(MicroED). Nat Methods 16(5):369–379.
https://doi.org/10.1038/s41592-019-0395x
3. Wolff AM, Young ID, Sierra RG, Brewster AS,
Martynowycz MW, Nango E, Sugahara M,
Nakane T, Ito K, Aquila A, Bhowmick A, Biel
JT, Carbajo S, Cohen AE, Cortez S,
Gonzalez A, Hino T, Im D, Koralek JD,
Kubo M, Lazarou TS, Nomura T, Owada S,
Samelson AJ, Tanaka T, Tanaka R, Thompson
EM, van den Bedem H, Woldeyes RA,
Yumoto F, Zhao W, Tono K, Boutet S,
Iwata S, Gonen T, Sauter NK, Fraser JS,
Thompson MC (2020) Comparing serial
X-ray crystallography and microcrystal electron
diffraction (MicroED) as methods for routine
structure determination from small macromolecular crystals. IUCrJ 7(Pt 2):306–323.
https://doi.org/10.1107/
S205225252000072X
4. Zatsepin NA, Li C, Colasurd P, Nannenga BL
(2019) The complementarity of serial femtosecond crystallography and MicroED for structure determination from microcrystals. Curr
Opin Struct Biol 58:286–293. https://doi.
org/10.1016/j.sbi.2019.06.004
5. Purdy MD, Shi D, Chrustowicz J, Hattne J,
Gonen T, Yeager M (2018) MicroED structures of HIV-1 Gag CTD-SP1 reveal binding
interactions with the maturation inhibitor bevirimat.
Proc
Natl
Acad
Sci
115
(52):13258–13263.
https://doi.org/10.
1073/pnas.1806806115
6. de la Cruz MJ, Hattne J, Shi D, Seidler P,
Rodriguez J, Reyes FE, Sawaya MR,
Cascio D, Weiss SC, Kim SK, Hinck CS,
Hinck AP, Calero G, Eisenberg D, Gonen T
(2017) Atomic-resolution structures from
fragmented protein crystals with the cryoEM
method MicroED. Nat Methods 14
(4):399–402.
https://doi.org/10.1038/
nmeth.4178
7. Nannenga BL, Shi D, Leslie AG, Gonen T
(2014) High-resolution structure determination by continuous-rotation data collection in
MicroED. Nat Methods 11(9):927–930.
https://doi.org/10.1038/nmeth.3043
8. Nannenga BL, Shi D, Hattne J, Reyes FE,
Gonen T (2014) Structure of catalase determined by MicroED. elife 3:e03600. https://
doi.org/10.7554/eLife.03600
9. Shi D, Nannenga BL, Iadanza MG, Gonen T
(2013) Three-dimensional electron crystallography of protein microcrystals. elife 2:e01345.
https://doi.org/10.7554/eLife.01345
10. Xu H, Lebrette H, Clabbers MTB, Zhao J,
Griese JJ, Zou X, Ho ¨gbom M (2019) Solving
a new R2lox protein structure by microcrystal
electron diffraction. Sci Adv 5(8):eaax4621.
https://doi.org/10.1126/sciadv.aax4621
11. Warmack RA, Boyer DR, Zee C-T, Richards
LS, Sawaya MR, Cascio D, Gonen T, Eisenberg
DS, Clarke SG (2019) Structure of amyloid-β
(20-34) with Alzheimer’s-associated isomerization at Asp23 reveals a distinct protofilament
interface. Nat Commun 10(1):3357. https://
doi.org/10.1038/s41467-019-11183-z
12. Krotee P, Griner SL, Sawaya MR, Cascio D,
Rodriguez JA, Shi D, Philipp S, Murray K,
Saelices L, Lee J, Seidler P, Glabe CG,
Jiang L, Gonen T, Eisenberg DS (2018) Common fibrillar spines of amyloid-beta and human
islet amyloid polypeptide revealed by microelectron diffraction and structure-based inhibitors. J Biol Chem 293(8):2888–2902. https://
doi.org/10.1074/jbc.M117.806109
13. Gallagher-Jones M, Glynn C, Boyer DR, Martynowycz MW, Hernandez E, Miao J, Zee CT,
Novikova IV, Goldschmidt L, McFarlane HT,
Helguera GF, Evans JE, Sawaya MR, Cascio D,
Eisenberg DS, Gonen T, Rodriguez JA (2018)
Sub-angstrom cryo-EM structure of a prion
protofibril reveals a polar clasp. Nat Struct
Mol Biol 25(2):131–134. https://doi.org/
10.1038/s41594-017-0018-0
14. Krotee P, Rodriguez JA, Sawaya MR, Cascio D,
Reyes FE, Shi D, Hattne J, Nannenga BL,
Oskarsson ME, Philipp S, Griner S, Jiang L,
Glabe CG, Westermark GT, Gonen T, Eisenberg DS (2017) Atomic structures of fibrillar
segments of hIAPP suggest tightly mated betasheets are important for cytotoxicity. elife 6:
e19273.
https://doi.org/10.7554/eLife.
19273
15. Sawaya MR, Rodriguez J, Cascio D, Collazo
MJ, Shi D, Reyes FE, Hattne J, Gonen T,
Eisenberg DS (2016) Ab initio structure determination from prion nanocrystals at atomic
resolution by MicroED. Proc Natl Acad Sci U
S A 113(40):11232–11236. https://doi.org/
10.1073/pnas.1606287113
16. Rodriguez JA, Ivanova MI, Sawaya MR,
Cascio D, Reyes FE, Shi D, Sangwan S,
Guenther EL, Johnson LM, Zhang M,
Jiang L, Arbing MA, Nannenga BL, Hattne J,
Whitelegge J, Brewster AS, Messerschmidt M,
Boutet S, Sauter NK, Gonen T, Eisenberg DS
MicroED Sample Preparation for Proteins
295
1. Nannenga BL (2020) MicroED methodology
and development. Struct Dyn 7(1):014304.
https://doi.org/10.1063/1.5128226
2. Nannenga BL, Gonen T (2019) The cryo-EM
method microcrystal electron diffraction
(MicroED). Nat Methods 16(5):369–379.
https://doi.org/10.1038/s41592-019-0395x
3. Wolff AM, Young ID, Sierra RG, Brewster AS,
Martynowycz MW, Nango E, Sugahara M,
Nakane T, Ito K, Aquila A, Bhowmick A, Biel
JT, Carbajo S, Cohen AE, Cortez S,
Gonzalez A, Hino T, Im D, Koralek JD,
Kubo M, Lazarou TS, Nomura T, Owada S,
Samelson AJ, Tanaka T, Tanaka R, Thompson
EM, van den Bedem H, Woldeyes RA,
Yumoto F, Zhao W, Tono K, Boutet S,
Iwata S, Gonen T, Sauter NK, Fraser JS,
Thompson MC (2020) Comparing serial
X-ray crystallography and microcrystal electron
diffraction (MicroED) as methods for routine
structure determination from small macromolecular crystals. IUCrJ 7(Pt 2):306–323.
https://doi.org/10.1107/
S205225252000072X
4. Zatsepin NA, Li C, Colasurd P, Nannenga BL
(2019) The complementarity of serial femtosecond crystallography and MicroED for structure determination from microcrystals. Curr
Opin Struct Biol 58:286–293. https://doi.
org/10.1016/j.sbi.2019.06.004
5. Purdy MD, Shi D, Chrustowicz J, Hattne J,
Gonen T, Yeager M (2018) MicroED structures of HIV-1 Gag CTD-SP1 reveal binding
interactions with the maturation inhibitor bevirimat.
Proc
Natl
Acad
Sci
115
(52):13258–13263.
https://doi.org/10.
1073/pnas.1806806115
6. de la Cruz MJ, Hattne J, Shi D, Seidler P,
Rodriguez J, Reyes FE, Sawaya MR,
Cascio D, Weiss SC, Kim SK, Hinck CS,
Hinck AP, Calero G, Eisenberg D, Gonen T
(2017) Atomic-resolution structures from
fragmented protein crystals with the cryoEM
method MicroED. Nat Methods 14
(4):399–402.
https://doi.org/10.1038/
nmeth.4178
7. Nannenga BL, Shi D, Leslie AG, Gonen T
(2014) High-resolution structure determination by continuous-rotation data collection in
MicroED. Nat Methods 11(9):927–930.
https://doi.org/10.1038/nmeth.3043
8. Nannenga BL, Shi D, Hattne J, Reyes FE,
Gonen T (2014) Structure of catalase determined by MicroED. elife 3:e03600. https://
doi.org/10.7554/eLife.03600
9. Shi D, Nannenga BL, Iadanza MG, Gonen T
(2013) Three-dimensional electron crystallography of protein microcrystals. elife 2:e01345.
https://doi.org/10.7554/eLife.01345
10. Xu H, Lebrette H, Clabbers MTB, Zhao J,
Griese JJ, Zou X, Ho ¨gbom M (2019) Solving
a new R2lox protein structure by microcrystal
electron diffraction. Sci Adv 5(8):eaax4621.
https://doi.org/10.1126/sciadv.aax4621
11. Warmack RA, Boyer DR, Zee C-T, Richards
LS, Sawaya MR, Cascio D, Gonen T, Eisenberg
DS, Clarke SG (2019) Structure of amyloid-β
(20-34) with Alzheimer’s-associated isomerization at Asp23 reveals a distinct protofilament
interface. Nat Commun 10(1):3357. https://
doi.org/10.1038/s41467-019-11183-z
12. Krotee P, Griner SL, Sawaya MR, Cascio D,
Rodriguez JA, Shi D, Philipp S, Murray K,
Saelices L, Lee J, Seidler P, Glabe CG,
Jiang L, Gonen T, Eisenberg DS (2018) Common fibrillar spines of amyloid-beta and human
islet amyloid polypeptide revealed by microelectron diffraction and structure-based inhibitors. J Biol Chem 293(8):2888–2902. https://
doi.org/10.1074/jbc.M117.806109
13. Gallagher-Jones M, Glynn C, Boyer DR, Martynowycz MW, Hernandez E, Miao J, Zee CT,
Novikova IV, Goldschmidt L, McFarlane HT,
Helguera GF, Evans JE, Sawaya MR, Cascio D,
Eisenberg DS, Gonen T, Rodriguez JA (2018)
Sub-angstrom cryo-EM structure of a prion
protofibril reveals a polar clasp. Nat Struct
Mol Biol 25(2):131–134. https://doi.org/
10.1038/s41594-017-0018-0
14. Krotee P, Rodriguez JA, Sawaya MR, Cascio D,
Reyes FE, Shi D, Hattne J, Nannenga BL,
Oskarsson ME, Philipp S, Griner S, Jiang L,
Glabe CG, Westermark GT, Gonen T, Eisenberg DS (2017) Atomic structures of fibrillar
segments of hIAPP suggest tightly mated betasheets are important for cytotoxicity. elife 6:
e19273.
https://doi.org/10.7554/eLife.
19273
15. Sawaya MR, Rodriguez J, Cascio D, Collazo
MJ, Shi D, Reyes FE, Hattne J, Gonen T,
Eisenberg DS (2016) Ab initio structure determination from prion nanocrystals at atomic
resolution by MicroED. Proc Natl Acad Sci U
S A 113(40):11232–11236. https://doi.org/
10.1073/pnas.1606287113
16. Rodriguez JA, Ivanova MI, Sawaya MR,
Cascio D, Reyes FE, Shi D, Sangwan S,
Guenther EL, Johnson LM, Zhang M,
Jiang L, Arbing MA, Nannenga BL, Hattne J,
Whitelegge J, Brewster AS, Messerschmidt M,
Boutet S, Sauter NK, Gonen T, Eisenberg DS
MicroED Sample Preparation for Proteins
295