arXiv subjects

Constantin R. Krüger

Publications and source records attributed to Constantin R. Krüger.

4 recordsLinked to original sources

arXiv2026

An Instrument for Physical Vapor Deposition onto Cryo-EM Samples for Microsecond Time-Resolved Cryo-EM

Laser flash melting and revitrification experiments have recently improved the time resolution of cryo-electron microscopy (cryo-EM) to the microsecond timescale, making it fast enough to observe many of the protein motions that are associated with function. The technique has also opened up a new dimension for cryo-EM sample preparation, making it possible to deposit compounds onto a cryo-EM sample while it is frozen, so that upon flash melting, the embedded particles experience an altered environment. For example, we have recently shown that depositing ultrathin silicon dioxide membranes onto a cryo-EM sample causes particles to detach from the interface upon flash melting, removing preferred particle orientation. These experiments also point towards a new strategy for initiating protein dynamics in time resolved experiments by depositing reagents, which will then mix with the sample upon flash melting. Here, we describe an apparatus for physical vapor deposition of compounds onto cryo-EM samples, detailing its design and operation. As a demonstration, we determine that the minimum thickness of silicon dioxide sealing membranes in a laser flash melting experiment is just over two monolayers. We propose that our design can form the basis for an integrated platform for microsecond time-resolved cryo-EM experiments.

physics.ins-det↗

arXiv2024

Shaped Laser Pulses for Microsecond Time-Resolved Cryo-EM: Outrunning Crystallization During Flash Melting

Water vitrifies if cooled at rates above $3 \cdot 10^5$ K/s. Surprisingly, this process cannot simply be reversed by heating the resulting amorphous ice at a similar rate. Instead, we have recently shown that the sample transiently crystallizes even if the heating rate is more than one order of magnitude higher. This may present an issue for microsecond time-resolved cryo-electron microscopy experiments, in which vitreous ice samples are briefly flash melted with a laser pulse, since transient crystallization could potentially alter the dynamics of the embedded proteins. Here, we demonstrate how shaped microsecond laser pulses can be used to increase the heating rate and outrun crystallization during flash melting of amorphous solid water (ASW) samples. We use time-resolved electron diffraction experiments to determine that the critical heating rate is about $10^8$ K/s, more than two orders of magnitude higher than the critical cooling rate. Our experiments add to the toolbox of the emerging field of microsecond time-resolved cryo-electron microscopy by demonstrating a straightforward approach for avoiding crystallization during laser melting and for achieving significantly higher heating rates, which paves the way for nanosecond time-resolved experiments.

physics.chem-ph↗

arXiv2023

Flash Melting Amorphous Ice

Water can be vitrified if it is cooled at rates exceeding $3*10^5$ K/s. This makes it possible to outrun crystallization in so-called no man's land, a range of deeply supercooled temperatures where water crystallizes rapidly. One would naively assume that the process can simply be reversed by heating the resulting amorphous ice at a similar rate. We demonstrate that this is not the case. When amorphous ice samples are flash melted with a microsecond laser pulse, time-resolved electron diffraction reveals that the sample transiently crystallizes despite a heating rate of more than $5*10^6$ K/s, demonstrating that the critical heating rate for outrunning crystallization is significantly higher than the critical cooling rate during vitrification. Moreover, we observe different crystallization kinetics for amorphous solid water (ASW) and hyperquenched glassy water (HGW), which suggests that the supercooled liquids formed during laser heating transiently retain distinct non-equilibrium structures that are associated with different nucleation rates. These experiments open up new avenues for elucidating the crystallization mechanism of water and studying its dynamics in no man's land. They also add important mechanistic details to the laser melting and revitrification process that is integral to the emerging field of microsecond time-resolved cryo-electron microscopy.

physics.chem-ph↗

arXiv2022

Electron Diffraction of Water in No Man's Land

A generally accepted understanding of the anomalous properties of water will only emerge if it becomes possible to systematically characterize water in the deeply supercooled regime, from where the anomalies appear to emanate. This has largely remained elusive because water crystallizes rapidly between 160 K and 232 K. Here, we present an experimental approach to rapidly prepare deeply supercooled water at a well-defined temperature and probe it with electron diffraction before crystallization occurs. We show that as water is cooled from room temperature to cryogenic temperature, its structure evolves smoothly, approaching that of amorphous ice just below 200 K. Our experiments narrow down the range of possible explanations of the origin for the water anomalies and open up new avenues for studying supercooled water.

physics.chem-ph↗