Time-resolved cryoEM using Spotiton

Dandey, V. P.; Budell, W. C.; Wei, H.; Bobe, D.; Maruthi, K.; Kopylov, M.; Eng, E. T.; Kahn, P. A.; Hinshaw, J. E.; Kundu, N.; Nimigean, C. M.; Fan, C.; Sukomon, N.; Darst, S.; Saecker, R.; Chen, J.; Malone, B.; Potter, C. S.; Carragher, B.

doi:10.1101/2020.04.20.050518

bioRxiv · 10.1101/2020.04.20.050518

Time-resolved cryoEM using Spotiton

Dandey, V. P.·Budell, W. C.·Wei, H.·Bobe, D.·Maruthi, K.·Kopylov, M.·Eng, E. T.·Kahn, P. A.·Hinshaw, J. E.·Kundu, N.·Nimigean, C. M.·Fan, C.·Sukomon, N.·Darst, S.·Saecker, R.·Chen, J.·Malone, B.·Potter, C. S.·Carragher, B.

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Abstract

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, on the order of ~100 ms later. We demonstrate the utility of this approach for four biological systems where short-lived states are of high interest.

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Dandey, V. P., Budell, W. C., Wei, H., Bobe, D., Maruthi, K., Kopylov, M., Eng, E. T., Kahn, P. A., Hinshaw, J. E., Kundu, N., Nimigean, C. M., Fan, C., Sukomon, N., Darst, S., Saecker, R., Chen, J., Malone, B., Potter, C. S., Carragher, B.. 2020-04-21. Time-resolved cryoEM using Spotiton. https://doi.org/10.1101/2020.04.20.050518

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