Abstract
X-ray lasers offer new capabilities in understanding the structure of biological systems, complex materials and matter under extreme conditions1,2,3,4. Very short and extremely bright, coherent X-ray pulses can be used to outrun key damage processes and obtain a single diffraction pattern from a large macromolecule, a virus or a cell before the sample explodes and turns into plasma1. The continuous diffraction pattern of non-crystalline objects permits oversampling and direct phase retrieval2. Here we show that high-quality diffraction data can be obtained with a single X-ray pulse from a non-crystalline biological sample, a single mimivirus particle, which was injected into the pulsed beam of a hard-X-ray free-electron laser, the Linac Coherent Light Source5. Calculations indicate that the energy deposited into the virus by the pulse heated the particle to over 100,000 K after the pulse had left the sample. The reconstructed exit wavefront (image) yielded 32-nm full-period resolution in a single exposure and showed no measurable damage. The reconstruction indicates inhomogeneous arrangement of dense material inside the virion. We expect that significantly higher resolutions will be achieved in such experiments with shorter and brighter photon pulses focused to a smaller area. The resolution in such experiments can be further extended for samples available in multiple identical copies.
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Acknowledgements
This work was supported by the following agencies: the Swedish Research Councils; the Swedish Foundation for International Cooperation in Research and Higher Education; Stiftelsen Olle Engkvist Byggmästare; the Swedish University of Agricultural Sciences; the Helmholtz Association (VH-VI-302); the DFG Cluster of Excellence at the Munich Centre for Advanced Photonics; the Centre National de la Recherche Scientifique; Agence Nationale de la Recherche (ANR-BLAN08-0089); the Hamburg Ministry of Science and Research and Joachim Herz Stiftung, as part of the Hamburg Initiative for Excellence in Research (LEXI); the Hamburg School for Structure and Dynamics; the Max Planck Society, the US National Science Foundation (grants MCB 0919195 and MCB-1021557); and the US Department of Energy, through the PULSE Institute. Portions of this research were carried out at the Linac Coherent Light Source, a National User Facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. We are grateful to B. Hedman and N. Timneanu for their help and to the scientific and technical staff of the LCLS for their outstanding facility and support.
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J.H. and H.N.C. conceived the experiment. C.A., V.S., J.-M.C., M.S., O.J., A. Rocker, I.A. and D.O. prepared and characterized the samples. J.D.B., C.B. and K.O.H. created the LCLS beamline. J.H., H.N.C., J.S., L.G., A.B., N.C., A.A., A.M., J.K., K.O.H. and S.P.H.-R. developed the imaging concept. S.W.E., R.H., D. Rolles, A. Rudenko, C.S., L.F., N.K., P.H., B.R., B.E., A.H., Ch.R., D.P., G.W., L.S., G.H., H. Gorke, J.U., I.S., S.H., G.S., F.S., H.S., K.-U.K., R.A., C.-D.S., F.K., M. Bott, S. Schorb, D. Rupp, M.A., T.G., H.H., L.G., G.P., H. Graafsma and B.N. designed and set up the CAMP instrument and/or developed and operated the pnCCD detectors. S. Bajt and M. Barthelmess coordinated instrumentation, worked on engineering and prepared filters and calibration samples. J.K., S.P.H.-R., A.B., H.N.C., J.S., A.M. and N.C. characterized the focus. B.I., D.W., M.S., M.M.S. and J.H. built the sample injector. D.P.D., J.H., M.S., D.W., U.W., R.K., M. Hunter, R.B.D. and J.C.H.S. designed and built the nanospray nebulizer. J.H., M.S., M.M.S., F.R.N.C.M., J.A., A.A.M., A. Rocker, M.J.B., C.Y.H., R.G.S., S. Boutet, I.A., O.J., D.S., A.B., J.S., D.P.D., A.A., M.L., J.K., T.A.W., A.M., R.L.S., L.L., M. Barthelmess, J.C.H.S., P.F., I.S., U.W., R.K., M. Hunter, R.B.D., M.F., G.W., M. Bott, S.W.E., B.E., L.F., R.H., N.K., L.L., D. Rupp, B.R., A. Rudenko, R.L.S., L.S., I.S., C.S., J.U. and H.N.C. characterized the imaging apparatus and carried out the experiment. T.E., F.R.N.C.M., M. Hantke, M.M.S., A.B., T.A.W., A.M., A.A., S.M., M.M. and J.H. analysed the data. T.E., F.R.N.C.M., A.B. and S.M. performed image reconstructions. J.H. and T.E. wrote the manuscript with input from all authors.
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Seibert, M., Ekeberg, T., Maia, F. et al. Single mimivirus particles intercepted and imaged with an X-ray laser. Nature 470, 78–81 (2011). https://doi.org/10.1038/nature09748
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DOI: https://doi.org/10.1038/nature09748
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