Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• eLife, 2022-01, Vol.11

Ort / Verlag

England: eLife Sciences Publications Ltd

Erscheinungsjahr

2022

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.