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Microsecond melting and revitrification of cryo samples

The dynamics of proteins that are associated with their function typically occur on the microsecond timescale, orders of magnitude faster than the time resolution of cryo-electron microscopy. We have recently introduced a novel approach to time-resolved cryo-electron microscopy that affords microsec...

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Autores principales: Voss, Jonathan M., Harder, Oliver F., Olshin, Pavel K., Drabbels, Marcel, Lorenz, Ulrich J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Crystallographic Association 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8550802/
https://www.ncbi.nlm.nih.gov/pubmed/34734102
http://dx.doi.org/10.1063/4.0000129
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author Voss, Jonathan M.
Harder, Oliver F.
Olshin, Pavel K.
Drabbels, Marcel
Lorenz, Ulrich J.
author_facet Voss, Jonathan M.
Harder, Oliver F.
Olshin, Pavel K.
Drabbels, Marcel
Lorenz, Ulrich J.
author_sort Voss, Jonathan M.
collection PubMed
description The dynamics of proteins that are associated with their function typically occur on the microsecond timescale, orders of magnitude faster than the time resolution of cryo-electron microscopy. We have recently introduced a novel approach to time-resolved cryo-electron microscopy that affords microsecond time resolution. It involves melting a cryo sample with a heating laser, so as to allow dynamics of the proteins to briefly occur in the liquid phase. When the laser is turned off, the sample rapidly revitrifies, trapping the particles in their transient configurations. Precise control of the temperature evolution of the sample is crucial for such an approach to succeed. Here, we provide a detailed characterization of the heat transfer occurring under laser irradiation as well as the associated phase behavior of the cryo sample. While areas close to the laser focus undergo melting and revitrification, surrounding regions crystallize. In situ observations of these phase changes therefore provide a convenient approach for assessing the temperature reached in each melting and revitrification experiment and for adjusting the heating laser power on the fly.
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spelling pubmed-85508022021-11-02 Microsecond melting and revitrification of cryo samples Voss, Jonathan M. Harder, Oliver F. Olshin, Pavel K. Drabbels, Marcel Lorenz, Ulrich J. Struct Dyn ARTICLES The dynamics of proteins that are associated with their function typically occur on the microsecond timescale, orders of magnitude faster than the time resolution of cryo-electron microscopy. We have recently introduced a novel approach to time-resolved cryo-electron microscopy that affords microsecond time resolution. It involves melting a cryo sample with a heating laser, so as to allow dynamics of the proteins to briefly occur in the liquid phase. When the laser is turned off, the sample rapidly revitrifies, trapping the particles in their transient configurations. Precise control of the temperature evolution of the sample is crucial for such an approach to succeed. Here, we provide a detailed characterization of the heat transfer occurring under laser irradiation as well as the associated phase behavior of the cryo sample. While areas close to the laser focus undergo melting and revitrification, surrounding regions crystallize. In situ observations of these phase changes therefore provide a convenient approach for assessing the temperature reached in each melting and revitrification experiment and for adjusting the heating laser power on the fly. American Crystallographic Association 2021-10-26 /pmc/articles/PMC8550802/ /pubmed/34734102 http://dx.doi.org/10.1063/4.0000129 Text en © 2021 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle ARTICLES
Voss, Jonathan M.
Harder, Oliver F.
Olshin, Pavel K.
Drabbels, Marcel
Lorenz, Ulrich J.
Microsecond melting and revitrification of cryo samples
title Microsecond melting and revitrification of cryo samples
title_full Microsecond melting and revitrification of cryo samples
title_fullStr Microsecond melting and revitrification of cryo samples
title_full_unstemmed Microsecond melting and revitrification of cryo samples
title_short Microsecond melting and revitrification of cryo samples
title_sort microsecond melting and revitrification of cryo samples
topic ARTICLES
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8550802/
https://www.ncbi.nlm.nih.gov/pubmed/34734102
http://dx.doi.org/10.1063/4.0000129
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