Cryo-EM structures from sub-nl volumes using pin-printing and jet vitrification

Raimond B. G. Ravelli (), Frank J. T. Nijpels, Rene J. M. Henderikx, Giulia Weissenberger, Sanne Thewessem, Abril Gijsbers, Bart W. A. M. M. Beulen, Carmen López-Iglesias and Peter J. Peters ()
Additional contact information
Raimond B. G. Ravelli: Maastricht University
Frank J. T. Nijpels: Maastricht University
Rene J. M. Henderikx: Maastricht University
Giulia Weissenberger: Maastricht University
Sanne Thewessem: Maastricht University
Abril Gijsbers: Maastricht University
Bart W. A. M. M. Beulen: CryoSol-World
Carmen López-Iglesias: Maastricht University
Peter J. Peters: Maastricht University

Nature Communications, 2020, vol. 11, issue 1, 1-9

Abstract: Abstract The increasing demand for cryo-electron microscopy (cryo-EM) reveals drawbacks in current sample preparation protocols, such as sample waste and lack of reproducibility. Here, we present several technical developments that provide efficient sample preparation for cryo-EM studies. Pin printing substantially reduces sample waste by depositing only a sub-nanoliter volume of sample on the carrier surface. Sample evaporation is mitigated by dewpoint control feedback loops. The deposited sample is vitrified by jets of cryogen followed by submersion into a cryogen bath. Because the cryogen jets cool the sample from the center, premounted autogrids can be used and loaded directly into automated cryo-EMs. We integrated these steps into a single device, named VitroJet. The device’s performance was validated by resolving four standard proteins (apoferritin, GroEL, worm hemoglobin, beta-galactosidase) to ~3 Å resolution using a 200-kV electron microscope. The VitroJet offers a promising solution for improved automated sample preparation in cryo-EM studies.

Date: 2020
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-020-16392-5 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-16392-5

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-16392-5

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().