- Time-resolved cryo-EM using Spotiton.
Time-resolved cryo-EM using Spotiton.
Nature methods (2020-08-12)
Venkata P Dandey, William C Budell, Hui Wei, Daija Bobe, Kashyap Maruthi, Mykhailo Kopylov, Edward T Eng, Peter A Kahn, Jenny E Hinshaw, Nidhi Kundu, Crina M Nimigean, Chen Fan, Nattakan Sukomon, Seth A Darst, Ruth M Saecker, James Chen, Brandon Malone, Clinton S Potter, Bridget Carragher
PMID32778833
要旨
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.