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Cryo-EM structures from sub-nl volumes using pin-printing and jet vitrification
Raimond B G Ravelli1, Frank J T Nijpels2,3, Rene J M Henderikx2,3
1The Maastricht Multimodal Molecular Imaging Institute (M4i), Division of Nanoscopy, Maastricht University, Maastricht, Netherlands. rbg.ravelli@maastrichtuniversity.nl.
Nature Communications
|May 24, 2020
Summary
New automated sample preparation for cryo-electron microscopy (cryo-EM) reduces waste and improves reproducibility. The VitroJet device enables high-resolution protein structure determination with cryo-EM.
Area of Science:
- Structural Biology
- Biophysics
- Biochemistry
Background:
- Current cryo-electron microscopy (cryo-EM) sample preparation methods face challenges with reproducibility and sample waste.
- High demand for cryo-EM necessitates improved, efficient, and automated sample preparation techniques.
Purpose of the Study:
- To present technical advancements for efficient cryo-EM sample preparation.
- To introduce an integrated device, VitroJet, for automated sample vitrification.
Main Methods:
- Pin printing technology for sub-nanoliter sample deposition, minimizing waste.
- Dewpoint control feedback loops to prevent sample evaporation.
- Cryogen jet and bath vitrification enabling direct use of autogrids in automated cryo-EM workflows.
Main Results:
- The VitroJet device successfully prepared samples for cryo-EM analysis.
- Four standard proteins (apoferritin, GroEL, worm hemoglobin, beta-galactosidase) were resolved to approximately 3 Å resolution.
- High-resolution data was obtained using a 200-kV electron microscope.
Conclusions:
- The VitroJet device provides an efficient and reproducible method for automated cryo-EM sample preparation.
- This technology addresses key limitations in current cryo-EM workflows.
- The VitroJet shows significant promise for advancing structural biology research through improved cryo-EM accessibility.
