We are pleased to announce that our manuscript, “An integrated workflow for structural virology with a 100 keV electron microscope,” is now available on bioRxiv:
In this work, we present a complete and practical workflow for applying cryo-electron microscopy (cryo-EM) to structural virology using a 100 keV electron microscope – an instrument that can be easily integrated into a biochemistry laboratory and provides significantly greater accessibility than the high-end 200 keV and 300 keV systems traditionally required. The manuscript details optimized strategies for sample preparation, data acquisition, and image processing, demonstrating that high-quality structural information on viral specimens can be obtained with lower-voltage microscopes.
As owners of an in-house 100 keV Tundra microscope (Thermo Fisher Scientific), our lab has been able to rapidly iterate on sample optimization and data collection without relying on external facilities. This direct access has enabled efficient troubleshooting, streamlined workflows, and consistent data quality, allowing us to obtain high-resolution structures. Our experience highlights the practical advantages of local, lower-voltage cryo-EM for day-to-day structural virology research.
We have successfully determined structures of several important viral glycoproteins, including coronavirus spike proteins, HIV envelope trimers, and Henipavirus fusion (F) proteins, all using a 100 keV microscope. Beyond single-particle analysis, the 100 keV platform also enabled detailed visualization of supramolecular assemblies such as nanoparticles, liposomes, virus-like particles (VLPs), and intact viruses. These results underscore the versatility of lower-voltage cryo-EM for both high-resolution protein structure determination and the structural characterization of complex, heterogeneous biological assemblies.
