Time-resolved cryoEM using Spotiton

We present an approach for preparing cryoEM grids to study short-lived molecular states. Using piezo electric dispensing, two independent streams of ~50 pL sample drops are deposited within 10 ms of each other onto a nanowire EM grid surface, and the mixing reaction stops when the grid is vitrified...

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Veröffentlicht in:Nature methods 2020-08, Vol.17 (9), p.897-900
Hauptverfasser: Dandey, Venkata P., Budell, William C., Wei, Hui, Bobe, Daija, Maruthi, Kashyap, Kopylov, Mykhailo, Eng, Edward T., Kahn, Peter A., Hinshaw, Jenny E., Kundu, Nidhi, Nimigean, Crina M., Fan, Chen, Sukomon, Nattakan, Darst, Seth A., Saecker, Ruth M., Chen, James, Malone, Brandon, Potter, Clinton S., Carragher, Bridget
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Sprache:eng
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Zusammenfassung:We present an approach for preparing cryoEM grids to study short-lived molecular states. Using piezo electric dispensing, two independent streams of ~50 pL sample drops are deposited within 10 ms of each other onto a nanowire EM grid surface, 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.
ISSN:1548-7091
1548-7105
DOI:10.1038/s41592-020-0925-6