Fast and Accurate Defocus Modulation for Improved Tunability of Cryo-EM Experiments

Overview

Journal IUCrJ

Date 2020 May 21

PMID 32431839

Citations 2

Authors

Radostin Danev

Hirofumi Iijima

Mizuki Matsuzaki

Sohei Motoki

Affiliations

Soon will be listed here.

Abstract

Current data collection strategies in electron cryo-microscopy (cryo-EM) record multiframe movies with static optical settings. This limits the number of adjustable parameters that can be used to optimize the experiment. Here, a method for fast and accurate defocus (FADE) modulation during movie acquisition is proposed. It uses the objective lens aperture as an electrostatic pole that locally modifies the electron beam potential. The beam potential variation is converted to defocus change by the typically undesired chromatic aberration of the objective lens. The simplicity, electrostatic principle and low electrical impedance of the device allow fast switching speeds that will enable per-frame defocus modulation of cryo-EM movies. Researchers will be able to define custom defocus 'recipes' and tailor the experiment for optimal information extraction from the sample. The FADE method could help to convert the microscope into a more dynamic and flexible optical platform that delivers better performance in cryo-EM single-particle analysis and electron cryo-tomography.

Citing Articles

Cryogenic electron ptychographic single particle analysis with wide bandwidth information transfer.

Pei X, Zhou L, Huang C, Boyce M, Kim J, Liberti E Nat Commun. 2023; 14(1):3027.

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CryoEM single particle reconstruction with a complex-valued particle stack.

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