Femtosecond X-ray Fourier holography imaging of free-flying nanoparticles


Ultrafast X-ray imaging on individual fragile specimens such as aerosols1, metastable particles2, superfluid quantum systems3 and live biospecimens4 provides high-resolution information that is inaccessible with conventional imaging techniques. Coherent X-ray diffractive imaging, however, suffers from intrinsic loss of phase, and therefore structure recovery is often complicated and not always uniquely defined4,5. Here, we introduce the method of in-flight holography, where we use nanoclusters as reference X-ray scatterers to encode relative phase information into diffraction patterns of a virus. The resulting hologram contains an unambiguous three-dimensional map of a virus and two nanoclusters with the highest lateral resolution so far achieved via single shot X-ray holography. Our approach unlocks the benefits of holography for ultrafast X-ray imaging of nanoscale, non-periodic systems and paves the way to direct observation of complex electron dynamics down to the attosecond timescale.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: Principles of in-flight holography.
Fig. 2: Experimental setup.
Fig. 3: Hologram and reconstructions.
Fig. 4: 3D reconstruction.
Fig. 5: Holograms of deformed specimen recorded with small reference clusters.


  1. 1.

    Loh, N. D. et al. Fractal morphology, imaging and mass spectrometry of single aerosol particles in flight. Nature 486, 513–517 (2012).

    ADS  Article  Google Scholar 

  2. 2.

    Barke, I. et al. The 3D-architecture of individual free silver nanoparticles captured by X-ray scattering. Nat. Commun. 6, 6187 (2015).

    Article  Google Scholar 

  3. 3.

    Gomez, L. F. et al. Shapes and vorticities of superfluid helium nanodroplets. Science 345, 906–909 (2014).

    ADS  Article  Google Scholar 

  4. 4.

    van der Schot, G. et al. Imaging single cells in a beam of live cyanobacteria with an X-ray laser. Nat. Commun. 6, 5704 (2015).

    Article  Google Scholar 

  5. 5.

    Seibert, M. M. et al. Single mimivirus particles intercepted and imaged with an X-ray laser. Nature 470, 78–81 (2011).

    ADS  Article  Google Scholar 

  6. 6.

    Neutze, R., Wouts, R., van der Spoel, D., Weckert, E. & Hajdu, J. Potential for biomolecular imaging with femtosecond X-ray pulses. Nature 406, 752–757 (2000).

    ADS  Article  Google Scholar 

  7. 7.

    Chapman, H. N. et al. Femtosecond X-ray protein nanocrystallography. Nature 470, 73–77 (2011).

    ADS  Article  Google Scholar 

  8. 8.

    Bostedt, C. et al. Ultrafast X-ray scattering of Xenon nanoparticles: imaging transient states of matter. Phys. Rev. Lett. 108, 093401 (2012).

    ADS  Article  Google Scholar 

  9. 9.

    Gorkhover, T. et al. Femtosecond and nanometre visualization of structural dynamics in superheated nanoparticles. Nat. Photon. 10, 93–97 (2016).

    ADS  Article  Google Scholar 

  10. 10.

    Miao, J, Sayre, D. & Chapman, H. Phase retrieval from the magnitude of the Fourier transforms of nonperiodic objects. J. Opt. Soc. Am A 15, 1662–1669 (1998).

    ADS  Article  Google Scholar 

  11. 11.

    Marchesini, S. et al. X-ray image reconstruction from a diffraction pattern alone. Phys. Rev. B 68, 140101(R) (2003).

    ADS  Article  Google Scholar 

  12. 12.

    Chapman, H. N. & Nugent, K. A. Coherent lensless X-ray imaging. Nat. Photon. 4, 833–839 (2010).

    ADS  Article  Google Scholar 

  13. 13.

    Gabor, D. et al. A new microscopic principle. Nature 161, 777–778 (1948).

    ADS  Article  Google Scholar 

  14. 14.

    Goodman, J. W. Introduction to Fourier Optics (Roberts and Company, Columbus, OH, USA, 2005).

  15. 15.

    Eisebitt, S. et al. Lensless imaging of magnetic nanostructures by X-ray. Nature 432, 885–888 (2004).

    ADS  Article  Google Scholar 

  16. 16.

    Schlotter, W. et al. Multiple reference Fourier transform holography with soft x rays. Appl. Phys. Lett. 89, 163112 (2006).

    ADS  Article  Google Scholar 

  17. 17.

    Marchesini, S. et al. Massively parallel X-ray holography. Nat. Photon. 2, 560–563 (2008).

    Article  Google Scholar 

  18. 18.

    Geilhufe, J. et al. Extracting depth information of 3-dimensional structures from a single-view X-ray Fourier-transform hologram. Opt. Express 22, 24959–24969 (2014).

    ADS  Article  Google Scholar 

  19. 19.

    Guehrs, E. et al. Wavefield back-propagation in high-resolution X-ray holography with a movable field of view. Opt. Express 18, 18922–18931 (2010).

    ADS  Article  Google Scholar 

  20. 20.

    Shintake, T. Possibility of single biomolecule imaging with coherent amplification of weak scattering X-ray photons. Phys. Rev. E 78, 041906 (2008).

  21. 21.

    Chamard, V. et al. Three-dimensional X-ray Fourier transform holography: the Bragg case. Phys. Rev. Lett. 104, 165501 (2010).

    ADS  Article  Google Scholar 

  22. 22.

    Xiao, C. et al. Structural studies of the giant Mimivirus. PLoS Biol. 7, 0958–0966 (2009).

    Article  Google Scholar 

  23. 23.

    Ferguson, K. R. et al. The atomic, molecular and optical science instrument at the linac coherent light source. J. Synchrot. Radiat. 22, 492–497 (2015).

    Article  Google Scholar 

  24. 24.

    Guinier, A. & Fournet, G. Small-Angle Scattering of X-Rays (Wiley, New York, USA, 1955).

  25. 25.

    Bostedt, C. et al. Clusters in intense FLASH pulses: ultrafast ionization dynamics and electron emission studied with spectroscopic and scattering techniques. J. Phys. B 12, 083004 (2010).

    Google Scholar 

  26. 26.

    Schropp, A. & Schroer, C. G. Dose requirements for resolving a given feature in an object by coherent X-ray diffraction imaging. New. J. Phys. 12, 035016 (2010).

    ADS  Article  Google Scholar 

  27. 27.

    Hantke, M. et al. High-throughput imaging of heterogeneous cell organelles with an X-ray laser. Nat. Photon. 8, 943–949 (2014).

    ADS  MathSciNet  Article  Google Scholar 

  28. 28.

    Rupp, D. et al. Coherent diffractive imaging of single helium nanodroplets with a high harmonic generation source. Nat. Commun. 8, 493 (2017).

    ADS  Article  Google Scholar 

  29. 29.

    Zherebtsov, S. et al. Controlled near-field enhanced electron acceleration from dielectric nanospheres with intense few-cycle laser fields. Nat. Phys. 7, 656–662 (2011).

    Article  Google Scholar 

  30. 30.

    Gorkhover, T. et al Nanoplasma dynamics of single large xenon clusters irradiated with superintense X-ray pulses from the Linac coherent light source free-electron laser. Phys. Rev. Lett 108, 245005 (2012).

    ADS  Article  Google Scholar 

  31. 31.

    Ekeberg, T. et al. Three-dimensional reconstruction of the giant Mimivirus particle with an X-ray free-electron laser. Phys. Rev. Lett. 114, 098102 (2015).

    ADS  Article  Google Scholar 

  32. 32.

    Emma, P. et al. First lasing and operation of an å ngstrom-wavelength free-electron laser. Nat. Photon. 4, 641–647 (2010).

    ADS  Article  Google Scholar 

  33. 33.

    Rupp, D. et al. Identification of twinned gas phase clusters by single-shot scattering with intense soft X-ray pulses. New. J. Phys. 14, 055016 (2012).

    ADS  Article  Google Scholar 

  34. 34.

    Rupp, D. et al. Generation and structure of extremely large clusters in pulsed jets. J. Chem. Phys. 141, 044306 (2014).

    ADS  Article  Google Scholar 

  35. 35.

    Gutt, C. et al. Single shot spatial and temporal coherence properties of the SLAC LINAC coherent light source in the hard X-ray regime. Phys. Rev. Lett. 108, 024801 (2012).

    ADS  Article  Google Scholar 

  36. 36.

    Andritschke, R., Hartner, G., Hartmann, R., Meidinger, N. & Struder, L. Data analysis for characterizing pnCCDs. In Nuclear Science Symposium Conference Record 2008 2166–2172 (IEEE, 2008).

  37. 37.

    Strüder, L. et al. Large-format, high-speed, X-ray pnCCDs combined with electron and ion imaging spectrometers in a multipurpose chamber for experiments at 4th generation light sources. Nucl. Instrum. Meth. A 614, 483–496 (2010).

    ADS  Article  Google Scholar 

  38. 38.

    Howells, M. et al. Toward a practical X-ray Fourier holography at high resolution. Nucl. Instrum. Meth. A 467, 864–867 (2001).

    ADS  Article  Google Scholar 

  39. 39.

    He, H. et al. Use of extended and prepared reference objects in experimental Fourier transform X-ray holography. Appl. Phys. Lett. 85, 2454–2456 (2004).

    ADS  Article  Google Scholar 

Download references


We would like to thank J. Geilhufe, E. Guehrs, A. Schropp and S. Eisebitt for many helpful discussions. T.G. acknowledges the P. Ewald fellowship from the Volkswagen Foundation and the Panofsky fellowship from SLAC National Accelerator Laboratory. We would like to thank J. Segal and A. Tomada from SLAC for providing high-resistivity Si wafers. Parts of this research were carried out at the Linac Coherent Light Source (LCLS) at the SLAC National Accelerator Laboratory. LCLS is an Office of Science User Facility operated for the US Department of Energy Office of Science by Stanford University. This work is supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, Division of Chemical Sciences, Geosciences, and Biosciences under contract no. DE-AC02-06CH11357 and contract no. DE-AC02-76SF00515. T.M. acknowledges financial support from BMBF (German Federal Ministry of Education and Research) projects 05K10KT2 and 05K13KT2 as well as DFG (German Research Foundation) BO3169/2-2. This work was supported by the Swedish Research Council, the Knut and Alice Wallenberg Foundation, the European Research Council, the Röntgen-Angström Cluster, ELI Extreme Light Infrastructure Phase 2 (CZ.02.1.01/0.0/0.0/15 008/0000162), ELIBIO (CZ.02.1.01/0.0/0.0/15 003/0000447) from the European Regional Development Fund, Material science and the Chalmers Area of Advance. F.R.N.C.M. acknowledges the Swedish Foundation for Strategic Research. Portions of this research were carried out at Brookhaven National Laboratory, operated under contract no. DE-SC0012704 from the US Department of Energy Office of Science. G.F. acknowledges the support of NKFIH K115504.

Author information




T.G. conceived the concept of 'in-flight' holography with two sources with support from C.B., T.M. and J.H. The project was led by T.G., C.B., F.R.N.C.M. and J.H. The experimental setup was designed and the experiment was performed by all authors. The bioparticle injector was operated by J.B., M.Sei. and K.M.. The cluster source was operated by K.F., M.B., C.B. and T.G. The biological samples were prepared by D.H., D.S.D.L., K.O. and M.Sve. The online and offline data analysis was carried out by F.M., T.E., M.F.H., B.J.D., C.N., A.M., G.v.S., M.B. and K.F. The images were analysed and processed by A.U. and T.G. The results were interpreted by A.U. and T.G. with input from C.B., G.F., T.M., F.R.N.C.M. and J.H. The manuscript was written by T.G. and A.U. with contributions from C.B., G.F., T.M., F.M. and J.H. and input from all authors.

Corresponding author

Correspondence to Tais Gorkhover.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Gorkhover, T., Ulmer, A., Ferguson, K. et al. Femtosecond X-ray Fourier holography imaging of free-flying nanoparticles. Nature Photon 12, 150–153 (2018). https://doi.org/10.1038/s41566-018-0110-y

Download citation

Further reading


Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing