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 Kimmel; Richard 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

doi:10.1038/nphoton.2014.270

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 journal › Article › peer-review

135 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 language	English (US)
Pages (from-to)	943-949
Number of pages	7
Journal	Nature Photonics
Volume	8
Issue number	12
DOIs	https://doi.org/10.1038/nphoton.2014.270
State	Published - Nov 27 2014
Externally published	Yes

ASJC Scopus subject areas

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

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10.1038/nphoton.2014.270

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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

Hantke, MF, Hasse, D, Maia, FRNC, Ekeberg, T, John, K, Svenda, M, Loh, ND, Martin, AV, Timneanu, N, Larsson, DSD, Van Der Schot, G, Carlsson, GH, Ingelman, M, Andreasson, J, Westphal, D, Liang, M, Stellato, F, Deponte, DP, Hartmann, R, Kimmel, N, Kirian, RA, Seibert, MM, Mühlig, K, Schorb, S, Ferguson, K, Bostedt, C, Carron, S, Bozek, JD, Rolles, D, Rudenko, A, Epp, S, Chapman, HN, Barty, A, Hajdu, J & Andersson, I 2014, 'High-throughput imaging of heterogeneous cell organelles with an X-ray laser', Nature Photonics, vol. 8, no. 12, pp. 943-949. https://doi.org/10.1038/nphoton.2014.270

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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.",

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AU - Hasse, Dirk

AU - Maia, Filipe R.N.C.

AU - Ekeberg, Tomas

AU - John, Katja

AU - Svenda, Martin

AU - Loh, N. Duane

AU - Martin, Andrew V.

AU - Timneanu, Nicusor

AU - Larsson, Daniel S.D.

AU - Van Der Schot, Gijs

AU - Carlsson, Gunilla H.

AU - Ingelman, Margareta

AU - Andreasson, Jakob

AU - Westphal, Daniel

AU - Liang, Mengning

AU - Stellato, Francesco

AU - Deponte, Daniel P.

AU - Hartmann, Robert

AU - Kimmel, Nils

AU - Kirian, Richard A.

AU - Seibert, M. Marvin

AU - Mühlig, Kerstin

AU - Schorb, Sebastian

AU - Ferguson, Ken

AU - Bostedt, Christoph

AU - Carron, Sebastian

AU - Bozek, John D.

AU - Rolles, Daniel

AU - Rudenko, Artem

AU - Epp, Sascha

AU - Chapman, Henry N.

AU - Barty, Anton

AU - Hajdu, Janos

AU - Andersson, Inger

PY - 2014/11/27

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

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