Rapid melting and revitrification as an approach to microsecond time-resolved cryo-electron microscopy

Proteins typically undergo dynamics on the microsecond to millisecond timescale, which is much faster than the time resolution of cryo-electron microscopy. Here, we propose a novel approach for microsecond time-resolved cryo-electron microscopy that involves 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 a liquid environment. After the laser pulse, the sample vitrifies, trapping particles in their transient configurations. As a proof of principle, we study the disassembly of particles after they incur structural damage.