Nanofluidic chips for cryo-EM structure determination from picoliter sample volumes

Stefan T. Huber, Edin Sarajlic, Roeland Huijink, Felix Weis, Wiel H. Evers, Arjen J. Jakobi*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

2 Citations (Scopus)
31 Downloads (Pure)

Abstract

Cryogenic electron microscopy has become an essential tool for structure determination of biological macromolecules. In practice, the difficulty to reliably prepare samples with uniform ice thickness still represents a barrier for routine high-resolution imaging and limits the current throughput of the technique. We show that a nanofluidic sample support with well-defined geometry can be used to prepare cryo-EM specimens with reproducible ice thickness from picoliter sample volumes. The sample solution is contained in electron-transparent nanochannels that provide uniform thickness gradients without further optimisation and eliminate the potentially destructive air-water interface. We demonstrate the possibility to perform high-resolution structure determination with three standard protein specimens. Nanofabricated sample supports bear potential to automate the cryo-EM workflow, and to explore new frontiers for cryo-EM applications such as time-resolved imaging and high-throughput screening.

Original languageEnglish
Article numbere72629
JournaleLife
Volume11
DOIs
Publication statusPublished - 2022

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