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TitleLaser phase plate improves structure determination of small proteins by cryo-EM.
Journal, issue, pagesScience, Page eaeh0665, Year 2026
Publish dateJun 11, 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 electron cryo-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 electron cryo-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. They have thus been unable to surpass results achieved by standard cryo-EM. Here, we show that the laser phase plate (LPP), installed in a custom, modern Titan Krios microscope, enhances the resolution in single-particle reconstruction of small proteins by improving specimen-motion correction, recovery of information from the early frames, as well as particle visualization, 3D 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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