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== Citation ==

Torino, Stefania / Dhurandhar, Mugdha / Stroobants, Annelore / Claessens, Raf / Efremov, Rouslan G.
Time-resolved cryo-EM using a combination of droplet microfluidics with on-demand jetting. 2023. Nature Methods, Vol. 20, No. 9, p. 1400-1408

== Abstract ==

Single-particle cryogenic electron microscopy (cryo-EM) allows
reconstruction of high-resolution structures of proteins in different
conformations. Protein function often involves transient functional
conformations, which can be resolved using time-resolved cryo-EM
(trEM). In trEM, reactions are arrested after a defined delay time by rapid
vitrification of protein solution on the EM grid. Despite the increasing
interest in trEM among the cryo-EM community, making trEM samples with
a time resolution below 100 ms remains challenging. Here we report the
design and the realization of a time-resolved cryo-plunger that combines
a droplet-based microfluidic mixer with a laser-induced generator of
microjets that allows rapid reaction initiation and plunge-freezing of
cryo-EM grids. Using this approach, a time resolution of 5 ms was achieved
and the protein density map was reconstructed to a resolution of 2.1 Å. trEM
experiments on GroEL:GroES chaperonin complex resolved the kinetics of
the complex formation and visualized putative short-lived conformations
of GroEL–ATP complex.

== Keywords ==

== Links ==

https://www.nature.com/articles/s41592-023-01967-z

== Related software ==

== Related methods ==

== Comments ==

2023Torino TimeResolved - Revision history

WikiSysop: Created page with "== Citation == Torino, Stefania / Dhurandhar, Mugdha / Stroobants, Annelore / Claessens, Raf / Efremov, Rouslan G. Time-resolved cryo-EM using a combination of droplet micro..."