Direct Measurement of the Critical Cooling Rate for the Vitrification of Water

TL;DR

This study precisely measures the critical cooling rate for vitrifying pure water using advanced cryo-EM techniques, providing a key reference for cryopreservation and cryo-EM experiments.

Contribution

It introduces a novel in situ, time-resolved cryo-EM method to directly measure the critical cooling rate of water, achieving high accuracy.

Findings

Critical cooling rate of water is 6.4×10^6 K/s.

Developed a microsecond laser pulse technique for cryo-EM.

Expanded the toolkit for microsecond time-resolved cryo-EM.

Abstract

The vitrification of aqueous solutions through rapid cooling is a remarkable achievement that launched the field of cryo-electron microscopy (cryo-EM) and has enabled the cryopreservation of biological specimens. For judging the feasibility of a vitrification experiment, the critical cooling rate of pure water is a frequently cited reference quantity. However, an accurate determination has remained elusive, with estimates varying by several orders of magnitude. Here, we employ in situ and time-resolved electron microscopy to obtain a precise measurement. We use shaped microsecond laser pulses to briefly melt an amorphous ice sample before flash freezing it with a variable, well-defined cooling rate. This allows us to directly measure the critical cooling rate of pure water, which we determine to be $6.4\cdot10^{6}$ K/s. Our experimental approach also expands the toolkit of microsecond time-resolved cryo-EM, an emerging technique, in which a cryo sample is similarly flash melted and revitrified with a laser pulse.