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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 7, Iss. 12 — Dec. 19, 2012

Internal structure of an intact Convallaria majalis pollen grain observed with X-ray Fresnel coherent diffractive imaging

Adrian P. Mancuso, Matthew R. Groves, Oleg E. Polozhentsev, Garth J. Williams, Ian McNulty, Claude Antony, Rachel Santarella-Mellwig, Aleksander V. Soldatov, Victor Lamzin, Andrew G. Peele, Keith A. Nugent, and Ivan A. Vartanyants  »View Author Affiliations

Optics Express, Vol. 20, Issue 24, pp. 26778-26785 (2012)

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We have applied Fresnel Coherent Diffractive Imaging (FCDI) to image an intact pollen grain from Convallaria majalis. This approach allows us to resolve internal structures without the requirement to chemically treat or slice the sample into thin sections. Coherent X-ray diffraction data from this pollen grain–composed of a hologram and higher resolution scattering information–was collected at a photon energy of 1820 eV and reconstructed using an iterative algorithm. A comparison with images recorded using transmission electron microscopy demonstrates that, while the resolution of these images is limited by the available flux and mechanical stability, we observed structures internal to the pollen grain–the intine/exine separations and pore dimensions–finer than 60 nm. The potential of this technique for further biological imaging applications is discussed.

© 2012 OSA

OCIS Codes
(000.4920) General : Other life sciences
(340.7440) X-ray optics : X-ray imaging
(340.7460) X-ray optics : X-ray microscopy

ToC Category:
X-ray Optics

Original Manuscript: September 10, 2012
Revised Manuscript: October 27, 2012
Manuscript Accepted: October 28, 2012
Published: November 13, 2012

Virtual Issues
Vol. 7, Iss. 12 Virtual Journal for Biomedical Optics

Adrian P. Mancuso, Matthew R. Groves, Oleg E. Polozhentsev, Garth J. Williams, Ian McNulty, Claude Antony, Rachel Santarella-Mellwig, Aleksander V. Soldatov, Victor Lamzin, Andrew G. Peele, Keith A. Nugent, and Ivan A. Vartanyants, "Internal structure of an intact Convallaria majalis pollen grain observed with X-ray Fresnel coherent diffractive imaging," Opt. Express 20, 26778-26785 (2012)

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  1. H. Breiteneder, K. Pettenburger, A. Bito, R. Valenta, D. Kraft, H. Rumpold, O. Scheiner, and M. Breitenbach, “The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene,” EMBO J.8(7), 1935–1938 (1989). [PubMed]
  2. H. Breiteneder, F. Ferreira, K. Hoffmann-Sommergruber, C. Ebner, M. Breitenbach, H. Rumpold, D. Kraft, and O. Scheiner, “Four recombinant isoforms of Cor a I, the major allergen of hazel pollen, show different IgE-binding properties,” Eur. J. Biochem.212(2), 355–362 (1993). [CrossRef] [PubMed]
  3. R. J. Scott and A. D. Stead, eds., Molecular and Cellular Aspects of Plant Reproduction (Cambridge University Press, 1994).
  4. L. Belin and J. R. Rowley, “Demonstration of birch pollen allergen from isolated pollen grains using immunofluorescence and a single radial immunodiffusion technique,” Int. Arch. Allergy Appl. Immunol.40(6), 754–769 (1971). [CrossRef] [PubMed]
  5. S. Vrtala, M. Grote, M. Duchêne, R. van Ree, D. Kraft, O. Scheiner, and R. Valenta, “Properties of tree and grass pollen allergens: reinvestigation of the linkage between solubility and allergenicity,” Int. Arch. Allergy Immunol.102(2), 160–169 (1993). [CrossRef] [PubMed]
  6. M. Grote, “Ultrastructural morphology and allergen detection in birch pollen after aqueous, anhydrous-liquid, and vapor fixation techniques,” Microsc. Res. Tech.21(3), 242–248 (1992). [CrossRef] [PubMed]
  7. M. Grote, “In situ localization of pollen allergens by immunogold electron microscopy: allergens at unexpected sites,” Int. Arch. Allergy Immunol.118(1), 1–6 (1999). [CrossRef] [PubMed]
  8. M. Grote, S. Vrtala, and R. Valenta, “Monitoring of two allergens, Bet v I and profilin, in dry and rehydrated birch pollen by immunogold electron microscopy and immunoblotting,” J. Histochem. Cytochem.41(5), 745–750 (1993). [CrossRef] [PubMed]
  9. M. Takahashi, J. W. Nowicke, G. L. Webster, S. S. Orli, and S. Yankowski, “Pollen morphology, exine structure, and systematics of Acalyphoideae (Euphorbiaceae), part 3. Tribes Epiprineae (Epiprinus, Symphyllia, Adenochlaena, Cleidiocarpon, Koilodepas, Cladogynos, Cephalocrotonopsis, Cephalocroton, Cephalomappa), Adelieae (Adelia, Crotonogynopsis, Enriquebeltrania, Lasiocroton, Leucocroton), Alchorneae (Orfilea, Alchornea, Coelebogyne, Aparisthmium, Bocquillonia, Conceveiba, Gavarretia), Acalypheae pro parte (Ricinus, Adriana, Mercurialis, Leidesia, Dysopsis, Wetria, Cleidion, Sampantaea, Macaranga),” Rev. Palaeobot. Palynol.110(1-2), 1–66 (2000). [CrossRef] [PubMed]
  10. L. Kovacik, J. M. Plitzko, M. Grote, and R. Reichelt, “Electron tomography of structures in the wall of hazel pollen grains,” J. Struct. Biol.166(3), 263–271 (2009). [CrossRef] [PubMed]
  11. D. Y. Parkinson, G. McDermott, L. D. Etkin, M. A. Le Gros, and C. A. Larabell, “Quantitative 3-D imaging of eukaryotic cells using soft X-ray tomography,” J. Struct. Biol.162(3), 380–386 (2008). [CrossRef] [PubMed]
  12. 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(2), 025506 (2006). [CrossRef] [PubMed]
  13. J. Miao, P. Charalambous, J. Kirz, and D. Sayre, “Extending the methodology of X-ray crystallography to allow imaging of micrometre-sized non-crystalline specimens,” Nature400(6742), 342–344 (1999). [CrossRef]
  14. J. Miao, H. N. Chapman, J. Kirz, D. Sayre, and K. O. Hodgson, “Taking X-ray diffraction to the limit: macromolecular structures from femtosecond X-ray pulses and diffraction microscopy of cells with synchrotron radiation,” Annu. Rev. Biophys. Biomol. Struct.33(1), 157–176 (2004). [CrossRef] [PubMed]
  15. K. A. Nugent, “Coherent methods in the X-ray sciences,” Adv. Phys.59(1), 1–99 (2010). [CrossRef]
  16. D. Shapiro, P. Thibault, T. Beetz, V. Elser, M. R. Howells, C. Jacobsen, J. Kirz, E. Lima, H. Miao, A. M. Neiman, and D. Sayre, “Biological imaging by soft x-ray diffraction microscopy,” Proc. Natl. Acad. Sci. U.S.A.102(43), 15343–15346 (2005). [CrossRef] [PubMed]
  17. J. Miao, K. O. Hodgson, T. Ishikawa, C. A. Larabell, M. A. LeGros, and Y. Nishino, “Imaging whole Escherichia coli bacteria by using single-particle x-ray diffraction,” Proc. Natl. Acad. Sci. U.S.A.100(1), 110–112 (2003). [CrossRef] [PubMed]
  18. C. Song, H. Jiang, A. P. Mancuso, B. Amirbekian, L. Peng, R. Sun, S. S. Shah, Z. H. Zhou, T. Ishikawa, and J. Miao, “Quantitative imaging of single, unstained viruses with coherent x Rays,” Phys. Rev. Lett.101(15), 158101 (2008). [CrossRef] [PubMed]
  19. X. Huang, J. Nelson, J. Kirz, E. Lima, S. Marchesini, H. Miao, A. M. Neiman, D. Shapiro, J. Steinbrener, A. Stewart, J. J. Turner, and C. Jacobsen, “Soft X-ray diffraction microscopy of a frozen hydrated yeast cell,” Phys. Rev. Lett.103(19), 198101 (2009). [CrossRef] [PubMed]
  20. E. Lima, L. Wiegart, P. Pernot, M. Howells, J. Timmins, F. Zontone, and A. Madsen, “Cryogenic x-ray diffraction microscopy for biological samples,” Phys. Rev. Lett.103(19), 198102 (2009). [CrossRef] [PubMed]
  21. Y. Nishino, Y. Takahashi, N. Imamoto, T. Ishikawa, and K. Maeshima, “Three-dimensional visualization of a human chromosome using coherent X-ray diffraction,” Phys. Rev. Lett.102(1), 018101 (2009). [CrossRef] [PubMed]
  22. A. P. Mancuso, T. Gorniak, F. Staier, O. M. Yefanov, R. Barth, C. Christophis, B. Reime, J. Gulden, A. Singer, M. E. Pettit, T. Nisius, T. Wilhein, C. Gutt, G. Grübel, N. Guerassimova, R. Treusch, J. Feldhaus, S. Eisebitt, E. Weckert, M. Grunze, A. Rosenhahn, and I. A. Vartanyants, “Coherent imaging of biological samples with femtosecond pulses at the free-electron laser FLASH,” New J. Phys.12(3), 035003 (2010). [CrossRef]
  23. A. P. Mancuso, O. M. Yefanov, and I. A. Vartanyants, “Coherent diffractive imaging of biological samples at synchrotron and free electron laser facilities,” J. Biotechnol.149(4), 229–237 (2010). [CrossRef] [PubMed]
  24. H. M. Quiney, K. A. Nugent, and A. G. Peele, “Iterative image reconstruction algorithms using wave-front intensity and phase variation,” Opt. Lett.30(13), 1638–1640 (2005). [CrossRef] [PubMed]
  25. J. M. Rodenburg, A. C. Hurst, A. G. Cullis, B. R. Dobson, F. Pfeiffer, O. Bunk, C. David, K. Jefimovs, and I. Johnson, “Hard-X-Ray Lensless Imaging of Extended Objects,” Phys. Rev. Lett.98(3), 034801 (2007). [CrossRef] [PubMed]
  26. P. Thibault, M. Dierolf, A. Menzel, O. Bunk, C. David, and F. Pfeiffer, “High-resolution scanning x-ray diffraction microscopy,” Science321(5887), 379–382 (2008). [CrossRef] [PubMed]
  27. K. Giewekemeyer, P. Thibault, S. Kalbfleisch, A. Beerlink, C. M. Kewish, M. Dierolf, F. Pfeiffer, and T. Salditt, “Quantitative biological imaging by ptychographic x-ray diffraction microscopy,” Proc. Natl. Acad. Sci. U.S.A.107(2), 529–534 (2010). [CrossRef] [PubMed]
  28. G. J. Williams, E. Hanssen, A. G. Peele, M. A. Pfeifer, J. Clark, B. Abbey, G. Cadenazzi, M. D. de Jonge, S. Vogt, L. Tilley, and K. A. Nugent, “High-resolution X-ray imaging of Plasmodium falciparum-infected red blood cells,” Cytometry A73(10), 949–957 (2008). [CrossRef] [PubMed]
  29. 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(6), 063823 (2009). [CrossRef]
  30. I. McNulty, A. Khounsary, Y. P. Feng, Y. Qian, J. Barraza, C. Benson, and D. Shu, “A beamline for 1–4 keV microscopy and coherence experiments at the Advanced Photon Source,” Rev. Sci. Instrum.67(9), 3372 (1996). [CrossRef]
  31. D. J. Vine, G. J. Williams, J. N. Clark, C. T. Putkunz, M. A. Pfeifer, D. Legnini, C. Roehrig, E. Wrobel, E. Huwald, G. van Riessen, B. Abbey, T. Beetz, J. Irwin, M. Feser, B. Hornberger, I. McNulty, K. A. Nugent, and A. G. Peele, “An in-vacuum x-ray diffraction microscope for use in the 0.7-2.9 keV range,” Rev. Sci. Instrum.83(3), 033703 (2012). [CrossRef] [PubMed]
  32. A. Thompson, D. Attwood, E. Gullikson, M. R. Howells, K. Kim, J. Kirz, J. Kortright, I. Lindau, P. Pianetta, and A. Robinson, “X-ray Data Booklet,” Lawrence Berkeley National Laboratory 1–38 (2001).
  33. G. J. Williams, H. M. Quiney, A. G. Peele, and K. A. Nugent, “Fresnel coherent diffractive imaging: treatment and analysis of data,” New J. Phys.12(3), 035020 (2010). [CrossRef]
  34. S. Marchesini, H. He, H. N. Chapman, S. Hau-Riege, A. Noy, M. R. Howells, U. Weierstall, and J. C. H. Spence, “X-ray image reconstruction from a diffraction pattern alone,” Phys. Rev. B68(14), 140101 (2003). [CrossRef]
  35. C. T. Putkunz, M. A. Pfeifer, A. G. Peele, G. J. Williams, H. M. Quiney, B. Abbey, K. A. Nugent, and I. McNulty, “Fresnel coherent diffraction tomography,” Opt. Express18(11), 11746–11753 (2010). [CrossRef] [PubMed]

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