Year: 2026
Type: preprint
Abstract: The extreme intensity of X-ray free-electron laser (XFEL) pulses has enabled imaging of radiation-sensitive samples, such as macromolecular microcrystals, beyond radiation damage limits. These sources have the potential to enable biomolecular single-particle imaging, similar to cryo-electron microscopy but without the need for cryo-fixation and with temporal resolution from femtoseconds to milliseconds. While this possibility was recognized before XFELs were built, the biological single-particle imaging work flow has previously only been demonstrated on large virus particles. Building on improvements in X-ray beam focusing, particle delivery, diffraction detection, and advanced analysis, here we demonstrate imaging of a single molecular complex, the giant-hemoglobin erythrocruorin (Ery) with X-ray laser pulses. Two-dimensional classes of diffraction patterns could be reconstructed to 15 Angstrom resolution, and 3D images to ~20 to 30 Angstrom, even though the 3D merged intensity in reciprocal space extended beyond 20 Angstrom. The resolution discrepancy is likely due to heterogeneity caused by gas-phase compaction of the complexes. With increased throughput, this approach could be used to reveal in-situ structural details during mass spectrometry studies of biomolecules, while improvements in sample delivery may provide ultrafast snapshot imaging of biological single-particles in their native-state beyond the limitations of radiation damage.
Source: PubMed
Authors: Xavier, Paul Lourdu, Morgan, Andrew J., Bielecki, Johan, Samanta, Amit K., Dold, Simon, Li, Chufeng, Gao, Xinyue, Passmore, Spencer K., Kurta, Ruslan P., Berberich, Tim B., De Santis, Emiliano, Zhang, Wenhui, E, Juncheng, Koua, Faisal H. M., Letrun, Romain, Gelisio, Luca, Turkot, Oleksii, Sobolev, Egor, Haas, Lukas V., Lübke, Jannik, He, Jingxuan, Memovich, Madeline B. F., Janson, Kevin, Lenzen, Stefanie, Rafie-Zinedine, Safi, Butola, Mansi, Yefanov, Oleksandr, Trost, Fabian, Kim, Chan, Popelar, Tomas, Sato, Tokushi, Vakili, Mohammad, Ovcharenko, Yevheniy, Boll, Rebecca, Usenko, Sergey, Baumann, Thomas M., De Fanis, Alberto, Senftleben, Björn, Porro, Matteo, Castoldi, Andrea, Dourki, Ibrahym, Danilevski, Cyril, Round, Ekaterina, Schmidt, Christina, Han, Huijong, Alvarez, Roberto, Chia, Ervin, Cazey, Cornelia, Seuring, Carolin, Sha, Ruojie, Azubel, Maia, Bushnell, David A., Boutet, Sébastien, Ovchinnikov, Sergey, Mancuso, Adrian P., Meyer, Michael, Bean, Richard, Barty, Anton, Maia, Filipe R. N. C., Kirian, Richard A., Grant, Thomas D., Loh, N. Duane, Caleman, Carl, Marklund, Erik G., Zatsepin, Nadia, Martin, Andrew V., Bajt, Saša, Kornberg, Roger D., Küpper, Jochen, Chapman, Henry N.
Institutions: The University of Melbourne, Deutsches Elektronen-Synchrotron DESY, Center for Free-Electron Laser Science, European X-Ray Free-Electron Laser, Uppsala University, Politecnico di Milano, University of Rome Tor Vergata, Istituto Nazionale di Fisica Nucleare, Roma Tor Vergata, Universität Hamburg, Hamburg Centre for Ultrafast Imaging, Ca' Foscari University of Venice, Istituto Nazionale di Fisica Nucleare, Sezione di Milano, Arizona State University, National University of Singapore, Centre for Structural Systems Biology, New York University, Stanford Medicine, Stanford University, SLAC National Accelerator Laboratory, Linac Coherent Light Source, Harvard University, Massachusetts Institute of Technology, Lawrence Berkeley National Laboratory, National Energy Research Scientific Computing Center, University at Buffalo, State University of New York, Swinburne University of Technology, RMIT University
Language: English
Subfield: Radiation
Field: Physics and Astronomy
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W7132087383