Laser phase plate for transmission electron microscopy

• Published in: Nature methods Vol. 16; no. 10; pp. 1016 - 1020
• Main Authors: Schwartz, Osip, Axelrod, Jeremy J., Campbell, Sara L., Turnbaugh, Carter, Glaeser, Robert M., Müller, Holger
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.10.2019; Nature Publishing Group
• Subjects: 631/1647/2258; 631/1647/245; 631/1647/328/1259; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Continuous wave lasers; Electron microscopy; Electrons; Electrostatic charging; Imaging; Laser beams; Lasers; Life Sciences; Light; Macromolecules; Methods; Microscopes; Microscopy, Electron, Transmission - methods; Phase contrast; Phase microscope; Phase plates; Proteomics; Static Electricity; Transmission electron microscopes; Transmission electron microscopy; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/s41592-019-0552-2

Transmission electron microscopy (TEM) of rapidly frozen biological specimens, or cryo-EM, would benefit from the development of a phase plate for in-focus phase contrast imaging. Several types of phase plates have been investigated, but rapid electrostatic charging of all such devices has hindered these efforts. Here, we demonstrate electron phase manipulation with a high-intensity continuous-wave laser beam, and use it as a phase plate for TEM. We demonstrate the laser phase plate by imaging an amorphous carbon film. The laser phase plate provides a stable and tunable phase shift without electrostatic charging or unwanted electron scattering. These results suggest the possibility for dose-efficient imaging of unstained biological macromolecules and cells. Phase plates improve contrast in cryo-electron microscopy, but suffer from electrostatic charging and electron scattering. A laser phase plate overcomes these problems and may improve imaging of biological specimens.