Time-resolved cryo-EM using Spotiton.

Dandey, Venkata P; Budell, William C; Wei, Hui; Bobe, Daija; Maruthi, Kashyap; Kopylov, Mykhailo; Eng, Edward T; Kahn, Peter A et al. · Nat Methods · 2020

basic_science · Level V

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Abstract

We present an approach for preparing cryo-electron microscopy (cryo-EM) grids to study short-lived molecular states. Using piezoelectric dispensing, two independent streams of ~50-pl droplets of sample are deposited within 10 ms of each other onto the surface of a nanowire EM grid, and the mixing reaction stops when the grid is vitrified in liquid ethane ~100 ms later. We demonstrate this approach for four biological systems where short-lived states are of high interest.

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