Microsecond melting and revitrification of cryo samples with a correlative light-electron microscopy approach

Author:

Bongiovanni Gabriele,Harder Oliver F.,Drabbels Marcel,Lorenz Ulrich J.

Abstract

We have recently introduced a novel approach to time-resolved cryo-electron microscopy (cryo-EM) that affords microsecond time resolution. It involves melting a cryo sample with a laser beam to allow dynamics of the embedded particles to occur. Once the laser beam is switched off, the sample revitrifies within just a few microseconds, trapping the particles in their transient configurations, which can subsequently be imaged to obtain a snap shot of the dynamics at this point in time. While we have previously performed such experiments with a modified transmission electron microscope, we here demonstrate a simpler implementation that uses an optical microscope. We believe that this will make our technique more easily accessible and hope that it will encourage other groups to apply microsecond time-resolved cryo-EM to study the fast dynamics of a variety of proteins.

Publisher

Frontiers Media SA

Subject

Biochemistry, Genetics and Molecular Biology (miscellaneous),Molecular Biology,Biochemistry

Cited by 12 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. Microsecond time-resolved cryo-electron microscopy;Current Opinion in Structural Biology;2024-08

2. Flash melting amorphous ice;The Journal of Chemical Physics;2024-05-09

3. Shaped Laser Pulses for Microsecond Time-Resolved Cryo-EM: Outrunning Crystallization during Flash Melting;The Journal of Physical Chemistry Letters;2024-04-11

4. Time resolved applications for Cryo-EM; approaches, challenges and future directions;Current Opinion in Structural Biology;2023-12

5. Fast viral dynamics revealed by microsecond time-resolved cryo-EM;Nature Communications;2023-09-13

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