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

Bongiovanni, Gabriele  
•
Harder, Oliver Florian  
•
Drabbels, Marcel  
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September 14, 2022

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.

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Name

2022.09.14.507962.full.pdf

Type

Preprint

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openaccess

License Condition

CC BY-NC-ND

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30.09 MB

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Adobe PDF

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abce744bd9f89e762f40213d2a19fc7b

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