Direct deposition on pre-cooled grids: a new approach to sub-millisecond cryoEM grid preparation.

Authors

David P Klebl, Robert W Kay, Sean McMillan, Frank Sobott, Nikil Kapur, Stephen P Muench

Published in

Acta crystallographica. Section D, Structural biology. Sep 01, 2026. Epub Sep 01, 2026.

Abstract

There has been a rapid expansion in new approaches to sample preparation for single-particle cryoEM. One drive has been around speeding up the process both for the application of time-resolved studies and in outrunning some of the negative effects which may come from the sample interacting with the air-water interface, such as degradation and preferred orientation. Here, we set out to significantly improve the speed of grid preparation by removing the plunging stage and directly spraying onto pre-cooled grids which are situated in the cryogen, allowing us to produce grids where two samples are mixed and frozen on the grid in less than 1 ms. We have previously shown a preliminary proof of principle with this new approach and here we provide a detailed description of the approach taken and the methodology behind it. This approach allows us to move beyond the millisecond timescale to open up more systems to time-resolved EM and potentially improve samples.

PMID:
42579337
Bibliographic data and abstract were imported from PubMed on 11 Aug 2026.

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