Abstract
X-ray tomography is an invaluable tool in biomedical imaging. It can deliver the three-dimensional internal structure of entire organisms as well as that of single cells, and even gives access to quantitative information, crucially important both for medical applications and for basic research1,2,3,4. Most frequently such information is based on X-ray attenuation. Phase contrast is sometimes used for improved visibility but remains significantly harder to quantify5,6. Here we describe an X-ray computed tomography technique that generates quantitative high-contrast three-dimensional electron density maps from phase contrast information without reverting to assumptions of a weak phase object or negligible absorption. This method uses a ptychographic coherent imaging approach to record tomographic data sets, exploiting both the high penetration power of hard X-rays and the high sensitivity of lensless imaging7,8,9. As an example, we present images of a bone sample in which structures on the 100 nm length scale such as the osteocyte lacunae and the interconnective canalicular network are clearly resolved. The recovered electron density map provides a contrast high enough to estimate nanoscale bone density variations of less than one per cent. We expect this high-resolution tomography technique to provide invaluable information for both the life and materials sciences.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Yin, G. et al. Energy-tunable transmission x-ray microscope for differential contrast imaging with near 60 nm resolution tomography. Appl. Phys. Lett. 88, 241115 (2006)
Parkinson, D. Y. et al. Quantitative 3-D imaging of eukaryotic cells using soft X-ray tomography. J. Struct. Biol. 162, 380–386 (2008)
Haddad, W. S. et al. Ultrahigh-resolution X-ray tomography. Science 266, 1213–1215 (1994)
Chu, Y. S. et al. Hard-x-ray microscopy with Fresnel zone plates reaches 40 nm Rayleigh resolution. Appl. Phys. Lett. 92, 103119 (2008)
Cloetens, P. et al. Phase objects in synchrotron radiation hard x-ray imaging. J. Phys. D 29, 133–146 (1996)
Wilkins, S. W. et al. Phase-contrast imaging using polychromatic hard X-rays. Nature 384, 335–338 (1996)
Nugent, K. Coherent methods in the X-ray sciences. Adv. Phys. 59, 1–99 (2010)
Rodenburg, J. M. Ptychography and related diffractive imaging methods. Adv. Imaging Electron Phys. 150, 87–184 (2008)
Thibault, P. et al. High-resolution scanning x-ray diffraction microscopy. Science 321, 379–382 (2008)
Davis, T. J. et al. Phase-contrast imaging of weakly absorbing materials using hard X-rays. Nature 373, 595–598 (1995)
Nugent, K. et al. Quantitative phase imaging using hard X rays. Phys. Rev. Lett. 77, 2961–2964 (1996)
Momose, A. et al. Phase-contrast X-ray computed tomography for observing biological soft tissues. Nature Med. 2, 473–475 (1996)
Pfeiffer, F. et al. Phase retrieval and differential phase-contrast imaging with low-brilliance X-ray sources. Nature Phys. 2, 258–261 (2006)
Faulkner, H. M. & Rodenburg, J. M. Movable aperture lensless transmission microscopy: a novel phase retrieval algorithm. Phys. Rev. Lett. 93, 023903 (2004)
Rodenburg, J. et al. Hard-x-ray lensless imaging of extended objects. Phys. Rev. Lett. 98, 034801 (2007)
Guizar-Sicairos, M. & Fienup, J. R. Phase retrieval with transverse translation diversity: a nonlinear optimization approach. Opt. Express 16, 7264–7278 (2008)
Giewekemeyer, K. et al. Quantitative biological imaging by ptychographic x-ray diffraction microscopy. Proc. Natl Acad. Sci. USA 107, 529–534 (2010)
Dierolf, M. et al. Ptychographic coherent diffractive imaging of weakly scattering specimens. N. J. Phys. 12, 035017 (2010)
Schropp, A. et al. Hard x-ray nanobeam characterization by coherent diffraction microscopy. Appl. Phys. Lett. 96, 091102 (2010)
Vine, D. J. et al. Ptychographic Fresnel coherent diffractive imaging. Phys. Rev. A 80, 063823 (2009)
Morrison, G. R. & Chapman, J. N. A comparison of three differential phase contrast systems suitable for use in STEM. Optik 64, 1–12 (1983)
Schneider, P. et al. Towards quantitative 3D imaging of the osteocyte lacuno-canalicular network. Bone 10.1016/j.bone.2010.07.026 (2010)
Kamioka, H. et al. A method for observing silver-stained osteocytes in situ in 3-m sections using ultra-high voltage electron microscopy tomography. Microsc. Microanal. 15, 377–383 (2009)
Hubbell, J. & Seltzer, M. Tables of X-ray Mass Attenuation Coefficients and Mass Energy-Absorption Coefficients. Version 1.4, Report NISTIR-5632 (National Institute of Standards and Technology, 1995) 〈http://physics.nist.gov/xaamdi〉.
Chapman, H. N. et al. High-resolution ab initio three-dimensional x-ray diffraction microscopy. J. Opt. Soc. Am. A 23, 1179–1200 (2006)
Nishino, Y. et al. Three-dimensional visualization of a human chromosome using coherent x-ray diffraction. Phys. Rev. Lett. 102, 018101 (2009)
Howells, M. et al. An assessment of the resolution limitation due to radiation-damage in X-ray diffraction microscopy. J. Electron Spectrosc. Relat. Phenom. 170, 4–12 (2009)
Kraft, P. et al. Performance of single-photon-counting PILATUS detector modules. J. Synchrotron Radiat. 16, 368–375 (2009)
Thibault, P. et al. Probe retrieval in ptychographic coherent diffractive imaging. Ultramicroscopy 109, 338–343 (2009)
Ghiglia, D. C. & Pritt, M. D. Two-Dimensional Phase Unwrapping: Theory, Algorithms And Software (Wiley, 1998)
Acknowledgements
We acknowledge technical support by X. Donath, FIB preparation of the specimen by P. Gasser and M. Meier and the assistance of W. Gutscher during the experiments. P.T., M.D. and F.P. acknowledge support through the DFG Cluster of Excellence “Munich-Centre for Advanced Photonics”.
Author information
Authors and Affiliations
Contributions
A.M., R.W., M.D., P.T., O.B., and F.P. conceived the experiment. R.W. prepared the sample. A.M., C.M.K., P.T., M.D., O.B. and F.P. carried out the experiment. P.T., M.D., C.M.K. and P.S. analysed the data. All authors discussed the results and contributed to the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
The file contains Supplementary Figure 1 and legend, legends for Supplementary Movies 1-6 and an additional reference. (PDF 274 kb)
Supplementary Movie 1
This movie shows reconstructed projections: amplitude (left), raw phase (centre), unwrapped phase (right) - see Supplementary Information file for full legend. (MOV 9440 kb)
Supplementary Movie 2
This movie shows sinograms calculated from amplitude and unwrapped phase projections - see Supplementary Information file for full legend. (MOV 9748 kb)
Supplementary Movie 3
This movie shows volume rendering of the reconstructed bone density - see Supplementary Information file for full legend. (MOV 9270 kb)
Supplementary Movie 4
This movie shows slices of tomographic volume perpendicular to the rotation axis - see Supplementary Information file for full legend. (MOV 9773 kb)
Supplementary Movie 5
This movie shows slices of tomographic volume in one of the directions parallel to the rotation axis - see Supplementary Information file for full legend. (MOV 11067 kb)
Supplementary Movie 6
This movie shows slices of tomographic volume parallel to the rotation axis and perpendicular to the slices in Movie 5 - see Supplementary Information file for full legend. (MOV 11063 kb)
Rights and permissions
About this article
Cite this article
Dierolf, M., Menzel, A., Thibault, P. et al. Ptychographic X-ray computed tomography at the nanoscale. Nature 467, 436–439 (2010). https://doi.org/10.1038/nature09419
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature09419
This article is cited by
-
Nanoscale imaging of Fe-rich inclusions in single-crystal zircon using X-ray ptycho-tomography
Scientific Reports (2024)
-
Three-dimensional nanoscale reduced-angle ptycho-tomographic imaging with deep learning (RAPID)
eLight (2023)
-
Accurate real space iterative reconstruction (RESIRE) algorithm for tomography
Scientific Reports (2023)
-
Deep learning at the edge enables real-time streaming ptychographic imaging
Nature Communications (2023)
-
4D nanoimaging of early age cement hydration
Nature Communications (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.