Rapid melting and revitrification as an approach to microsecond time-resolved cryo-electron microscopy
| Autor: | Oliver F. Harder, Jonathan M. Voss, Pavel K. Olshin, Marcel Drabbels, Ulrich J. Lorenz |
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| Rok vydání: | 2021 |
| Předmět: |
Materials science
Cryo-electron microscopy General Physics and Astronomy FOS: Physical sciences 02 engineering and technology Trapping 010402 general chemistry 01 natural sciences law.invention Quantitative Biology::Subcellular Processes law Microscopy Physics - Biological Physics Physical and Theoretical Chemistry Quantitative Biology::Biomolecules Millisecond business.industry Time resolution 021001 nanoscience & nanotechnology Laser 0104 chemical sciences Pulse (physics) Microsecond Biological Physics (physics.bio-ph) Optoelectronics 0210 nano-technology business |
| Zdroj: | Chemical Physics Letters |
| ISSN: | 0009-2614 |
| DOI: | 10.1016/j.cplett.2021.138812 |
| Popis: | Proteins typically undergo conformational dynamics on the microsecond to millisecond timescale as they perform their function, which is much faster than the time-resolution of cryo-electron microscopy and has thus prevented real-time observations. Here, we propose a novel approach for microsecond time-resolved cryo-electron microscopy that involves rapidly melting a cryo specimen in situ with a laser beam. The sample remains liquid for the duration of the laser pulse, offering a tunable time window in which the dynamics of embedded particles can be induced in their native liquid environment. After the laser pulse, the sample vitrifies in just a few microseconds, trapping particles in their transient configurations, so that they can subsequently be characterized with conventional cryo-electron microscopy. We demonstrate that our melting and revitrification approach is viable and affords microsecond time resolution. As a proof of principle, we study the disassembly of particles after they incur structural damage and trap them in partially unraveled configurations. |
| Databáze: | OpenAIRE |
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