High-throughput imaging of heterogeneous cell organelles with an X-ray laser

Max F. Hantke, Dirk Hasse, Filipe R.N.C. Maia, Tomas Ekeberg, Katja John, Martin Svenda, N. Duane Loh, Andrew V. Martin, Nicusor Timneanu, Daniel S.D. Larsson, Gijs Van Der Schot, Gunilla H. Carlsson, Margareta Ingelman, Jakob Andreasson, Daniel Westphal, Mengning Liang, Francesco Stellato, Daniel P. Deponte, Robert Hartmann, Nils KimmelRichard A. Kirian, M. Marvin Seibert, Kerstin Mühlig, Sebastian Schorb, Ken Ferguson, Christoph Bostedt, Sebastian Carron, John D. Bozek, Daniel Rolles, Artem Rudenko, Sascha Epp, Henry N. Chapman, Anton Barty, Janos Hajdu, Inger Andersson

Research output: Contribution to journalArticle

90 Scopus citations

Abstract

We overcome two of the most daunting challenges in single-particle diffractive imaging: collecting many high-quality diffraction patterns on a small amount of sample and separating components from mixed samples. We demonstrate this on carboxysomes, which are polyhedral cell organelles that vary in size and facilitate up to 40% of Earth's carbon fixation. A new aerosol sample-injector allowed us to record 70,000 low-noise diffraction patterns in 12...min with the Linac Coherent Light Source running at 120...Hz. We separate different structures directly from the diffraction data and show that the size distribution is preserved during sample delivery. We automate phase retrieval and avoid reconstruction artefacts caused by missing modes. We attain the highest-resolution reconstructions on the smallest single biological objects imaged with an X-ray laser to date. These advances lay the foundations for accurate, high-throughput structure determination by flash-diffractive imaging and offer a means to study structure and structural heterogeneity in biology and elsewhere.

Original languageEnglish (US)
Pages (from-to)943-949
Number of pages7
JournalNature Photonics
Volume8
Issue number12
DOIs
StatePublished - Nov 27 2014

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics

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    Hantke, M. F., Hasse, D., Maia, F. R. N. C., Ekeberg, T., John, K., Svenda, M., Loh, N. D., Martin, A. V., Timneanu, N., Larsson, D. S. D., Van Der Schot, G., Carlsson, G. H., Ingelman, M., Andreasson, J., Westphal, D., Liang, M., Stellato, F., Deponte, D. P., Hartmann, R., ... Andersson, I. (2014). High-throughput imaging of heterogeneous cell organelles with an X-ray laser. Nature Photonics, 8(12), 943-949. https://doi.org/10.1038/nphoton.2014.270