Regensburg 2025 – scientific programme
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BP: Fachverband Biologische Physik
BP 27: Focus Session Chemical Imaging for the Elucidation of Molecular Structure I (joint session O/BP)
BP 27.6: Talk
Thursday, March 20, 2025, 17:00–17:15, H24
Instrumentation for high-resolution biomolecule imaging enabled by electrospray ion beam deposition (ES-IBD) — •Lukas Eriksson1, Tim Esser1,2, and Stephan Rauschenbach1 — 1University of Oxford, Oxford, UK — 2Thermo Fisher Scientific, Eindhoven, Netherlands
Direct imaging of (bio-)molecules with cryogenic electron microscopy (cryo-EM) or scanning probe microscopy (SPM) is a powerful approach for elucidating molecular structure. However, sample preparation can be a major challenge: either very time- and resource-intensive or incompatible with the vacuum environment required by the imaging method.
Here, we explore preparative mass spectrometry as an alternative workflow towards structural elucidation of biomolecules. A novel, custom-built deposition stage extending a commercial mass spectrometer (Thermo Fisher Scientific Orbitrap UHMR) allows for the mass-filtered, soft-landed deposition of a wide mass range of target molecules (m = 100 to 106 Da) onto various surfaces, including cryo-EM grids and metal crystals for SPM. Successful deposition and subsequent imaging requires extensive control over conditions such as pressure, temperature, ion trajectories, sample surfaces, and sample transfer to obtain clean, chemically pure samples of the desired species in the right (i.e. native) configuration. The sample holder also enables controlled growth of ice layers for embedding deposited molecules, allowing high-resolution reconstructions of proteins from cryo-EM.
Keywords: Scanning Probe Microscopy (SPM); Cryo-EM; Mass Spectrometry; Instrumentation; Electrospray Ion Beam Deposition (ES-IBD)