Scanning X-ray fluorescence microscopy (XFM) is a particularly useful method for studying the spatial distribution of trace metals in biological samples. Here we demonstrate the utility of combining coherent diffractive imaging (CDI) with XFM for imaging biological samples to simultaneously produce high-resolution and high-contrast transmission images and quantitative elemental maps. The reconstructed transmission function yields morphological details which contextualise the elemental maps. We report enhancement of the spatial resolution in both the transmission and fluorescence images beyond that of the X-ray optics. The freshwater diatom Cyclotella meneghiniana was imaged to demonstrate the benefits of combining these techniques that have complementary contrast mechanisms.

© 2012 OSA

1. T. Paunesku, S. Vogt, J. Maser, B. Lai, and G. Woloschak, “X-ray fluorescence microprobe imaging in biology and medicine,” J. Cell Biochem.99, 1489–1502 (2006). [CrossRef] [PubMed]
2. B. S. Twining, S. B. Baines, N. S. Fisher, J. Maser, S. Vogt, C. Jacobsen, A. Tovar-Sanchez, and S. A. San̅udo-Wilhelmy, “Quantifying trace elements in individual aquatic protist cells with a synchrotron x-ray fluorescence microprobe,” Anal. Chem.75, 3806–3816 (2003). [CrossRef] [PubMed]
3. H. N. Chapman and K. A. Nugent, “Coherent lensless x-ray imaging,” Nat. Photonics4, 833–839 (2010). [CrossRef]
4. S. W. Hell, “Far-field optical nanoscopy,” Science316, 1153–1158 (2007). [CrossRef] [PubMed]
5. M. Bates, B. Huang, G. T. Dempsey, and X. Zhuang, “Multicolor super-resolution imaging with photo-switchable fluorescent probes,” Science317, 1749–1753 (2007). [CrossRef] [PubMed]
6. B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy,” Science319, 810–813 (2008). [CrossRef] [PubMed]
7. A. A. Snigirev, I. I. Snigreva, V. G. Kohn, S. Kuznetsov, and I. A. Schelokov, “On the possibilities of x-ray phase contrast microimaging by coherent high-energy synchrotron radiation,” Rev. Sci. Instrum.66, 5486–5492 (1995). [CrossRef]
8. B. Hornberger, M. D. de Jonge, M. Feser, P. Holl, C. Jacobsen, D. Legnini, P. Rehak, L. Struder, and S. Vogt, “Differential phase contrast with a segmented detector in a scanning x-ray microprobe,” J. Synch. Rad.15, 355–362 (2008). [CrossRef]
9. C. Holzner, M. Feser, S. Vogt, B. Hornberger, S. B. Baines, and C. Jacobsen, “Zernike phase contrast in scanning microscopy with x-rays,” Nat. Phys.6, 883–887 (2010). [CrossRef]
10. J. Miao, D. Sayre, and H. N. Chapman, “Phase retrieval from the magnitude of the Fourier transforms of nonperiodic objects,” J. Opt. Soc. Am. A15, 1662–1669 (1998). [CrossRef]
11. J. Miao, P. Charalambous, J. Kirz, and D. Sayre, “Extending the methodology of x-ray crystallography to micrometre-sized non-crystalline specimens,” Nature400, 342–344 (1999). [CrossRef]
12. I. K. Robinson, I. A. Vartanyants, G. J. Williams, M. A. Pfeifer, and J. A. Pitney, “Reconstruction of the shapes of gold nanocrystals using coherent x-ray diffraction,” Phys. Rev. Lett.87, 195505 (2001). [CrossRef] [PubMed]
13. G. J. Williams, H. M. Quiney, B. B. Dhal, C. Q. Tran, K. A. Nugent, A. G. Peele, D. Paterson, and M. D. de Jonge, “Fresnel coherent diffractive imaging,” Phys. Rev. Lett.97, 025506 (2006). [CrossRef] [PubMed]
14. K. A. Nugent, “Coherent methods in the x-ray sciences,” Adv. Phys.59, 1–99 (2010). [CrossRef]
15. H. M. L. Faulkner and J. M. Rodenburg, “Error tolerance of an iterative phase retrieval algorithm for movable illumination microscopy,” Ultramicroscopy103, 153–164 (2005). [CrossRef] [PubMed]
16. P. Thibault, M. Dierolf, A. Menzel, O. Bunk, C. David, and F. Pfeiffer, “High resolution scanning x-ray diffrction microscopy,” Science321, 379–382 (2008). [CrossRef] [PubMed]
17. D. J. Vine, G. J. Williams, B. Abbey, M. A. Pfeifer, J. N. Clark, M. D. De Jonge, I. McNulty, A. G. Peele, and K. A. Nugent, “Ptychographic Fresnel coherent diffractive imaging,” Phys. Rev. A80, 063823 (2009). [CrossRef]
18. Factorising the exit surface wave into the product of the incident beam multiplied by the sample transmission function requires assuming the sample is sufficiently thin for the projection approximation to hold [14, 34, 35].
19. P. Thibault, M. Dierolf, O. Bunk, A. Menzel, and F. Pfeiffer, “Probe retrieval in ptychographic coherent diffractive imaging,” Ultramicroscopy109, 338–343 (2009). [CrossRef] [PubMed]
20. L. Hare, “Aquatic insects and trace metals: Bioavailability, bioaccumulation, and toxicity,” Crc. Cr. Rev. Toxicol.22, 327–369 (1992). [CrossRef]
21. K. H. Thompson and C. Orvig, “Boon and bane of metal ions in medicine,” Science300, 936–939 (2003). [CrossRef] [PubMed]
22. A. Schropp, P. Boye, A. Goldschmidt, S. Hönig, R. Hoppe, J. Patommel, C. Rakete, D. Samberg, S. Stephan, S. Schröder, M. Burghammer, and C. Schroer, “Non-destructive and quantitative imaging of a nano-structured microchip by ptychographic hard x-ray scanning microscopy,” J. Microsc.241, 9–12 (2011). [CrossRef]
23. X. Llopart, M. Campbell, R. Dinapoli, D. San Segundo, and E. Pernigotti, “Medipix2: A 64-k pixel readout chip with 55 μm square elements working in single photon counting mode,” IEEE T. Nucl. Sci.49, 2279–2283 (2002). [CrossRef]
24. C. B. Field, M. J. Behrenfeld, J. T. Randerson, and P. Falkowski, “Primary production of the biosphere: Integrating terrestrial and oceanic components,” Science281, 237–240 (1998). [CrossRef] [PubMed]
25. R. A. Armstrong, C. Lee, J. I. Hedges, S. Honjo, and S. G. Wakeham, “A new, mechanistic model for organic carbon fluxes in the ocean based on the quantitative association of POC with ballast minerals,” Deep-Sea Res.Pt. 249, 219–236 (2002). [CrossRef]
26. K. O. Buesseler, “The decoupling of production and particulate export in the surface ocean,” Global Biogeochem. Cy.12, 297–310 (1998). [CrossRef]
27. J. Nagy and Z. Strakos, “Enforcing nonnegativity in image reconstruction algorithms,” in Proc. SPIE4121 (2000). [CrossRef]
28. C. Ryan, R. Kirkham, R. Hough, G. Moorhead, D. Siddons, M. de Jonge, D. Paterson, G. De Geronimo, D. Howard, and J. Cleverley, “Elemental x-ray imaging using the maia detector array: The benefits and challenges of large solid-angle,” Nucl. Instrum. Meth. A619, 37–43 (2010). [CrossRef]
29. W. Vernon, M. Allin, R. Hamlin, T. Hontz, D. Nguyen, F. Augustine, S. M. Gruner, N. H. Xuong, D. R. Schuette, M. W. Tate, and L. J. Koerner, “First results from the 128x128 pixel mixed-mode Si x-ray detector chip,” Proc. SPIE6706, 67060U (2007). [CrossRef]
30. M. D. de Jonge, C. Holzner, S. B. Baines, B. S. Twining, K. Ignatyev, J. Diaz, D. L. Howard, D. Legnini, A. Miceli, I. McNulty, C. J. Jacobsen, and S. Vogt, “Quantitative 3D elemental microtomography of cyclotella meneghiniana at 400-nm resolution,” P. Natl. Acad. Sci. USA107, 15676–15680 (2010). [CrossRef]
31. R. W. Gerchberg and W. O. Saxton, “A practical algorithm for the determination of phase from image and diffraction plane pictures,” Optik35, 237–246 (1972).
32. J. Fienup, T. Crimmins, and W. Holsztynski, “Reconstruction of the support of an object from the support of its autocorrelation,” J. Opt. Soc. Am. A72, 610–624 (1982). [CrossRef]
33. H. M. L. Faulkner and J. M. Rodenburg, “Movable aperture lens transmission microscopy: A novel phase retrieval algorithm,” Phys. Rev. Lett.93, 023903 (2004). [CrossRef] [PubMed]
34. J. M. Cowley, Diffraction Physics, 3rd ed. (North-Holland Publishing Company, Amsterdam, 1995).
35. D. M. Paganin, Coherent X-Ray Optics (Oxford University Press, New York, 2006). [CrossRef]

Adv. Phys.

• K. A. Nugent, “Coherent methods in the x-ray sciences,” Adv. Phys.59, 1–99 (2010). [CrossRef]

Anal. Chem.

• B. S. Twining, S. B. Baines, N. S. Fisher, J. Maser, S. Vogt, C. Jacobsen, A. Tovar-Sanchez, and S. A. San̅udo-Wilhelmy, “Quantifying trace elements in individual aquatic protist cells with a synchrotron x-ray fluorescence microprobe,” Anal. Chem.75, 3806–3816 (2003). [CrossRef] [PubMed]

Crc. Cr. Rev. Toxicol.

• L. Hare, “Aquatic insects and trace metals: Bioavailability, bioaccumulation, and toxicity,” Crc. Cr. Rev. Toxicol.22, 327–369 (1992). [CrossRef]

Deep-Sea Res.Pt. 2

• R. A. Armstrong, C. Lee, J. I. Hedges, S. Honjo, and S. G. Wakeham, “A new, mechanistic model for organic carbon fluxes in the ocean based on the quantitative association of POC with ballast minerals,” Deep-Sea Res.Pt. 249, 219–236 (2002). [CrossRef]

Global Biogeochem. Cy.

• K. O. Buesseler, “The decoupling of production and particulate export in the surface ocean,” Global Biogeochem. Cy.12, 297–310 (1998). [CrossRef]

IEEE T. Nucl. Sci.

• X. Llopart, M. Campbell, R. Dinapoli, D. San Segundo, and E. Pernigotti, “Medipix2: A 64-k pixel readout chip with 55 μm square elements working in single photon counting mode,” IEEE T. Nucl. Sci.49, 2279–2283 (2002). [CrossRef]

J. Cell Biochem.

• T. Paunesku, S. Vogt, J. Maser, B. Lai, and G. Woloschak, “X-ray fluorescence microprobe imaging in biology and medicine,” J. Cell Biochem.99, 1489–1502 (2006). [CrossRef] [PubMed]

J. Microsc.

• A. Schropp, P. Boye, A. Goldschmidt, S. Hönig, R. Hoppe, J. Patommel, C. Rakete, D. Samberg, S. Stephan, S. Schröder, M. Burghammer, and C. Schroer, “Non-destructive and quantitative imaging of a nano-structured microchip by ptychographic hard x-ray scanning microscopy,” J. Microsc.241, 9–12 (2011). [CrossRef]

J. Opt. Soc. Am. A

• J. Fienup, T. Crimmins, and W. Holsztynski, “Reconstruction of the support of an object from the support of its autocorrelation,” J. Opt. Soc. Am. A72, 610–624 (1982). [CrossRef]
• J. Miao, D. Sayre, and H. N. Chapman, “Phase retrieval from the magnitude of the Fourier transforms of nonperiodic objects,” J. Opt. Soc. Am. A15, 1662–1669 (1998). [CrossRef]

J. Synch. Rad.

• B. Hornberger, M. D. de Jonge, M. Feser, P. Holl, C. Jacobsen, D. Legnini, P. Rehak, L. Struder, and S. Vogt, “Differential phase contrast with a segmented detector in a scanning x-ray microprobe,” J. Synch. Rad.15, 355–362 (2008). [CrossRef]

Nat. Photonics

• H. N. Chapman and K. A. Nugent, “Coherent lensless x-ray imaging,” Nat. Photonics4, 833–839 (2010). [CrossRef]

Nat. Phys.

• C. Holzner, M. Feser, S. Vogt, B. Hornberger, S. B. Baines, and C. Jacobsen, “Zernike phase contrast in scanning microscopy with x-rays,” Nat. Phys.6, 883–887 (2010). [CrossRef]

Nature

• J. Miao, P. Charalambous, J. Kirz, and D. Sayre, “Extending the methodology of x-ray crystallography to micrometre-sized non-crystalline specimens,” Nature400, 342–344 (1999). [CrossRef]

Nucl. Instrum. Meth. A

• C. Ryan, R. Kirkham, R. Hough, G. Moorhead, D. Siddons, M. de Jonge, D. Paterson, G. De Geronimo, D. Howard, and J. Cleverley, “Elemental x-ray imaging using the maia detector array: The benefits and challenges of large solid-angle,” Nucl. Instrum. Meth. A619, 37–43 (2010). [CrossRef]

Optik

• R. W. Gerchberg and W. O. Saxton, “A practical algorithm for the determination of phase from image and diffraction plane pictures,” Optik35, 237–246 (1972).

P. Natl. Acad. Sci. USA

• M. D. de Jonge, C. Holzner, S. B. Baines, B. S. Twining, K. Ignatyev, J. Diaz, D. L. Howard, D. Legnini, A. Miceli, I. McNulty, C. J. Jacobsen, and S. Vogt, “Quantitative 3D elemental microtomography of cyclotella meneghiniana at 400-nm resolution,” P. Natl. Acad. Sci. USA107, 15676–15680 (2010). [CrossRef]

Phys. Rev. A

• D. J. Vine, G. J. Williams, B. Abbey, M. A. Pfeifer, J. N. Clark, M. D. De Jonge, I. McNulty, A. G. Peele, and K. A. Nugent, “Ptychographic Fresnel coherent diffractive imaging,” Phys. Rev. A80, 063823 (2009). [CrossRef]

Phys. Rev. Lett.

• I. K. Robinson, I. A. Vartanyants, G. J. Williams, M. A. Pfeifer, and J. A. Pitney, “Reconstruction of the shapes of gold nanocrystals using coherent x-ray diffraction,” Phys. Rev. Lett.87, 195505 (2001). [CrossRef] [PubMed]
• G. J. Williams, H. M. Quiney, B. B. Dhal, C. Q. Tran, K. A. Nugent, A. G. Peele, D. Paterson, and M. D. de Jonge, “Fresnel coherent diffractive imaging,” Phys. Rev. Lett.97, 025506 (2006). [CrossRef] [PubMed]
• H. M. L. Faulkner and J. M. Rodenburg, “Movable aperture lens transmission microscopy: A novel phase retrieval algorithm,” Phys. Rev. Lett.93, 023903 (2004). [CrossRef] [PubMed]

Proc. SPIE

• W. Vernon, M. Allin, R. Hamlin, T. Hontz, D. Nguyen, F. Augustine, S. M. Gruner, N. H. Xuong, D. R. Schuette, M. W. Tate, and L. J. Koerner, “First results from the 128x128 pixel mixed-mode Si x-ray detector chip,” Proc. SPIE6706, 67060U (2007). [CrossRef]
• J. Nagy and Z. Strakos, “Enforcing nonnegativity in image reconstruction algorithms,” in Proc. SPIE4121 (2000). [CrossRef]

Rev. Sci. Instrum.

• A. A. Snigirev, I. I. Snigreva, V. G. Kohn, S. Kuznetsov, and I. A. Schelokov, “On the possibilities of x-ray phase contrast microimaging by coherent high-energy synchrotron radiation,” Rev. Sci. Instrum.66, 5486–5492 (1995). [CrossRef]

Science

• S. W. Hell, “Far-field optical nanoscopy,” Science316, 1153–1158 (2007). [CrossRef] [PubMed]
• M. Bates, B. Huang, G. T. Dempsey, and X. Zhuang, “Multicolor super-resolution imaging with photo-switchable fluorescent probes,” Science317, 1749–1753 (2007). [CrossRef] [PubMed]
• B. Huang, W. Wang, M. Bates, and X. Zhuang, “Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy,” Science319, 810–813 (2008). [CrossRef] [PubMed]
• C. B. Field, M. J. Behrenfeld, J. T. Randerson, and P. Falkowski, “Primary production of the biosphere: Integrating terrestrial and oceanic components,” Science281, 237–240 (1998). [CrossRef] [PubMed]
• K. H. Thompson and C. Orvig, “Boon and bane of metal ions in medicine,” Science300, 936–939 (2003). [CrossRef] [PubMed]
• P. Thibault, M. Dierolf, A. Menzel, O. Bunk, C. David, and F. Pfeiffer, “High resolution scanning x-ray diffrction microscopy,” Science321, 379–382 (2008). [CrossRef] [PubMed]

Ultramicroscopy

• P. Thibault, M. Dierolf, O. Bunk, A. Menzel, and F. Pfeiffer, “Probe retrieval in ptychographic coherent diffractive imaging,” Ultramicroscopy109, 338–343 (2009). [CrossRef] [PubMed]
• H. M. L. Faulkner and J. M. Rodenburg, “Error tolerance of an iterative phase retrieval algorithm for movable illumination microscopy,” Ultramicroscopy103, 153–164 (2005). [CrossRef] [PubMed]

Other

• Factorising the exit surface wave into the product of the incident beam multiplied by the sample transmission function requires assuming the sample is sufficiently thin for the projection approximation to hold [14, 34, 35].
• J. M. Cowley, Diffraction Physics, 3rd ed. (North-Holland Publishing Company, Amsterdam, 1995).
• D. M. Paganin, Coherent X-Ray Optics (Oxford University Press, New York, 2006). [CrossRef]

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