Modular microfluidics enables kinetic insight from time-resolved cryo-EM.

ABSTRACT: Mechanistic understanding of biochemical reactions requires structural and kinetic characterization of the underlying chemical processes. However, no single experimental technique can provide this information in a broadly applicable manner and thus structural studies of static macromolecules are often complemented by biophysical analysis. Moreover, the common strategy of utilizing mutants or crosslinking probes to stabilize intermediates is prone to trapping off-pathway artefacts and precludes determining the order of molecular events. Here we report a time-resolved sample preparation method for cryo-electron microscopy (trEM) using a modular microfluidic device, featuring a 3D-mixing unit and variable delay lines that enables automated, fast, and blot-free sample vitrification. This appro

SUBMITTER: Maeots ME 

PROVIDER: S-EPMC7351747 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature