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TitleLaser phase plate improves structure determination of small proteins by cryo-EM.
Journal, issue, pagesScience, Vol. 393, Issue 6807, Page 195-199, Year 2026
Publish dateJul 9, 2026
AuthorsPetar N Petrov / Jessie T Zhang / Jonathan Remis / Jeremy J Axelrod / Hang Cheng / Eric S Cooper / Ian K Hicklin / Shahar Sandhaus / Cooper Schnurr / Robert M Glaeser / Holger Müller /
PubMed AbstractPhase 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 ...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.
External linksScience / PubMed:42275466
MethodsEM (single particle)
Resolution2.43 - 4.3 Å
Structure data

EMDB-76790: Rabbit Aldolase, Laser Phase Plate TEM, Laser-ON dataset 1 (A1)
Method: EM (single particle) / Resolution: 2.58 Å

EMDB-76791: Rabbit Aldolase, Laser Phase Plate TEM, Laser-OFF dataset 1 (A1)
Method: EM (single particle) / Resolution: 3.05 Å

EMDB-76792: Rabbit Aldolase, Laser Phase Plate TEM, Laser-ON dataset 2 (A2)
Method: EM (single particle) / Resolution: 2.43 Å

EMDB-76793: Rabbit Aldolase, Laser Phase Plate TEM, Laser-OFF dataset 2 (A2)
Method: EM (single particle) / Resolution: 2.46 Å

EMDB-76794: Rabbit Aldolase, Laser Phase Plate TEM, Laser-ON dataset 3 (A3)
Method: EM (single particle) / Resolution: 2.58 Å

EMDB-76797: Rabbit Aldolase, Laser Phase Plate TEM, Laser-OFF dataset 3 (A3)
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-76802: Human Hemoglobin, Laser Phase Plate TEM, Laser-ON dataset 1 (H1)
Method: EM (single particle) / Resolution: 3.04 Å

EMDB-76804: Human Hemoglobin, Laser Phase Plate TEM, Laser-OFF dataset 1 (H1)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-76805: Human Hemoglobin, Laser Phase Plate TEM, Laser-ON dataset 2 (H2)
Method: EM (single particle) / Resolution: 2.86 Å

EMDB-76806: Human Hemoglobin, Laser Phase Plate TEM, Laser-OFF dataset 2 (H2)
Method: EM (single particle) / Resolution: 3.34 Å

EMDB-76807: Human Hemoglobin, Laser Phase Plate TEM, Laser-ON dataset 3 (H3)
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-76809: Human Hemoglobin, Laser Phase Plate TEM, Laser-OFF dataset 3 (H3)
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-76810: Human Hemoglobin, Laser Phase Plate TEM, Laser-ON, Refined initial 3D class (IC-ON)
Method: EM (single particle) / Resolution: 3.64 Å

EMDB-76811: Human Hemoglobin, Laser Phase Plate TEM, Laser-OFF, Refined initial 3D class (IC-OFF)
Method: EM (single particle) / Resolution: 4.3 Å

Source
  • Oryctolagus cuniculus (rabbit)
  • Homo sapiens (human)

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