Laser phase plate improves structure determination of small proteins by cryo-EM
Abstract
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.
Article Details
Journal Info
Science
American Association for the Advancement of Science
Authors (11)
Petar N. Petrov
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Jessie T. Zhang
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Jonathan Remis
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Jeremy J. Axelrod
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Hang Cheng
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
Eric S. Cooper
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Ian K. Hicklin
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Shahar Sandhaus
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Cooper Schnurr
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.
Robert M. Glaeser
Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Holger Müller
Department of Physics, University of California, Berkeley, Berkeley, CA, USA.