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 Lopez-Iglesias, Peter J. Peters*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


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 similar to 3 angstrom resolution using a 200-kV electron microscope. The VitroJet offers a promising solution for improved automated sample preparation in cryo-EM studies. There is a need to further improve the automation of cryo-EM sample preparation to make it more easily accessible for non-specialists, reduce sample waste and increase reproducibility. Here, the authors present VitroJet, a single device, where sub-nl volumes of samples are deposited by pin printing thus eliminating the need for sample blotting, which is followed by jet vitrification, and they show that high-resolution structures can be obtained using four standard proteins.

Original languageEnglish
Article number2563
Number of pages9
JournalNature Communications
Issue number1
Publication statusPublished - 22 May 2020



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