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Automated cryo-EM sample preparation by pin-printing and jet vitrification

By 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, Peter J. Peters

Posted 28 May 2019
bioRxiv DOI: 10.1101/651208

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 controlled and 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 4 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.

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