---
OA_type: closed access
_id: '22365'
abstract:
- lang: eng
text: Phase plates can, in principle, overcome the poor image contrast in cryo–electron
microscopy (cryo-EM) and the resulting limits on the structural reconstruction
of small proteins. However, previous designs have been unstable and compromised
the high-resolution signal and have thus been unable to surpass results achieved
by standard cryo-EM. Here, we show that the laser phase plate (LPP), installed
in a modern, custom Titan Krios microscope, enhances the resolution in single-particle
reconstruction of small proteins by improving specimen-motion correction and recovery
of information from the early frames, as well as particle visualization, three-dimensional
classification, and alignment. These advances use standard defocus ranges and
reconstruction procedures but open the door to LPP-tailored protocols, offering
further improvements by leveraging the LPP demonstrated here.
acknowledgement: 'The authors thank O. Schwartz and S. Scheres for helpful remarks
and discussions; D. Agard, B. Carragher, C. Potter, and P. Olshin for close collaboration;
A. Singh, L. Maisenbacher, S. Strasser, and I. Pope for help with mirror inspection;
J. Fang, E. Nogales, and J. Hurley for sharing their lab space and assisting with
sample preparation; B. Buijsse, W. Hagen, B. Jiang, and T. Coyle at Thermo Fisher
Scientific for the design of the custom transfer optics and technical support; G.
Long and T. Gutierrez at the UC Berkeley Physics R&D Machine Shop for machining
cavity components and tooling. This work was supported by the following: Chan Zuckerberg
Initiative award numbers 2021-234606 and 2025-367757, National Institutes of Health
grant R01GM126011, Gordon and Betty Moore Foundation grant 9366, Lawrence Berkeley
National Laboratory Directed Research and Development Program grant 25-111, and
Cooperative Research and Development Agreement award AWD00004352 (to H.M.); National
Institutes of Health fellowship F32GM149186 (to P.N.P.).'
article_processing_charge: No
article_type: original
author:
- first_name: Petar N
full_name: Petrov, Petar N
id: b1d6732d-8cb6-11f0-baab-bd460ee3a287
last_name: Petrov
- first_name: Jessie T.
full_name: Zhang, Jessie T.
last_name: Zhang
- first_name: Jonathan
full_name: Remis, Jonathan
last_name: Remis
- first_name: Jeremy J.
full_name: Axelrod, Jeremy J.
last_name: Axelrod
- first_name: Hang
full_name: Cheng, Hang
last_name: Cheng
- first_name: Eric S.
full_name: Cooper, Eric S.
last_name: Cooper
- first_name: Ian K.
full_name: Hicklin, Ian K.
last_name: Hicklin
- first_name: Shahar
full_name: Sandhaus, Shahar
last_name: Sandhaus
- first_name: Cooper
full_name: Schnurr, Cooper
last_name: Schnurr
- first_name: Robert M.
full_name: Glaeser, Robert M.
last_name: Glaeser
- first_name: Holger
full_name: Müller, Holger
last_name: Müller
citation:
ama: Petrov PN, Zhang JT, Remis J, et al. Laser phase plate improves structure determination
of small proteins by cryo-EM. Science. 2026;393(6807):195-196. doi:10.1126/science.aeh0665
apa: Petrov, P. N., Zhang, J. T., Remis, J., Axelrod, J. J., Cheng, H., Cooper,
E. S., … Müller, H. (2026). Laser phase plate improves structure determination
of small proteins by cryo-EM. Science. AAAS. https://doi.org/10.1126/science.aeh0665
chicago: Petrov, Petar N, Jessie T. Zhang, Jonathan Remis, Jeremy J. Axelrod, Hang
Cheng, Eric S. Cooper, Ian K. Hicklin, et al. “Laser Phase Plate Improves Structure
Determination of Small Proteins by Cryo-EM.” Science. AAAS, 2026. https://doi.org/10.1126/science.aeh0665.
ieee: P. N. Petrov et al., “Laser phase plate improves structure determination
of small proteins by cryo-EM,” Science, vol. 393, no. 6807. AAAS, pp. 195–196,
2026.
ista: Petrov PN, Zhang JT, Remis J, Axelrod JJ, Cheng H, Cooper ES, Hicklin IK,
Sandhaus S, Schnurr C, Glaeser RM, Müller H. 2026. Laser phase plate improves
structure determination of small proteins by cryo-EM. Science. 393(6807), 195–196.
mla: Petrov, Petar N., et al. “Laser Phase Plate Improves Structure Determination
of Small Proteins by Cryo-EM.” Science, vol. 393, no. 6807, AAAS, 2026,
pp. 195–96, doi:10.1126/science.aeh0665.
short: P.N. Petrov, J.T. Zhang, J. Remis, J.J. Axelrod, H. Cheng, E.S. Cooper, I.K.
Hicklin, S. Sandhaus, C. Schnurr, R.M. Glaeser, H. Müller, Science 393 (2026)
195–196.
das_tickbox: '1'
dataavailabilitystatement: 'The datasets are publicly available in the Electron Microscopy
Public Image Archive [A1: EMPIAR-13528 (on), EMPIAR-13527 (off); A2: EMPIAR-13529
(on), EMPIAR-13526 (off); A3: EMPIAR-13530 (on), EMPIAR-13525 (off); H1: EMPIAR-13535
(on), EMPIAR-13533 (off); H2: EMPIAR-13534 (on), EMPIAR-13532 (off); H3: EMPIAR-13537
(on), EMPIAR-13531 (off)]. The final reconstructed maps are deposited in the Electron
Microscopy Data Bank [A1: EMD-76790 (on), EMD-76791 (off); A2: EMD-76792 (on), EMD-76793
(off); A3: EMD-76794 (on), EMD-76797 (off); H1: EMD-76802 (on), EMD-76804 (off);
H2: EMD-76805 (on), EMD-76806 (off); H3: EMD-76807 (on), EMD-76809 (off)]. The initial
structures in Fig. 3 are deposited at EMD-76810 (on) and EMD-76811 (off). Code for
converting EER movies to binned TIF format with proper accounting for electron dose
is deposited in Zenodo (47) and available on GitHub at https://github.com/matterwaves/eer2tiff/releases/tag/v0.1.0.
All specimen preparation materials are commercially available.'
date_created: 2026-07-19T22:01:46Z
date_published: 2026-07-09T00:00:00Z
date_updated: 2026-07-20T12:58:46Z
day: '09'
ddc:
- '570'
department:
- _id: MiLe
doi: 10.1126/science.aeh0665
external_id:
pmid:
- '42275466'
has_accepted_license: '1'
intvolume: ' 393'
issue: '6807'
language:
- iso: eng
month: '07'
oa_version: None
page: 195-196
pmid: 1
publication: Science
publication_identifier:
eissn:
- 1095-9203
issn:
- 0036-8075
publication_status: published
publisher: AAAS
quality_controlled: '1'
researchdata_availability: yes
scopus_import: '1'
status: public
supplementarymaterial: yes
title: Laser phase plate improves structure determination of small proteins by cryo-EM
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 393
year: '2026'
...