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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 5 — Feb. 27, 2012
  • pp: 5629–5635

Dynamic and high-resolution live cell imaging by direct electron beam excitation

Yasunori Nawa, Wataru Inami, Akito Chiba, Atsushi Ono, Atsuo Miyakawa, Yoshimasa Kawata, Sheng Lin, and Susumu Terakawa  »View Author Affiliations

Optics Express, Vol. 20, Issue 5, pp. 5629-5635 (2012)

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We propose a direct electron-beam excitation assisted optical microscope with a resolution of a few tens of nanometers and it can be applied for observation of dynamic movements of nanoparticles in liquid. The technique is also useful for live cell imaging under physiological conditions as well as observation of colloidal solution, microcrystal growth in solutions, etc. In the microscope, fluorescent materials are directly excited with a focused electron beam. The direct excitation with an electron beam yields high spatial resolution since the electron beam can be focused to a few tens of nanometers in the specimens. In order to demonstrate the potential of our proposed microscope, we observed the movements of fluorescent nanoparticles, which can be used for labelling specimens, in a water-based solution. We also demonstrated an observation result of living CHO cells.

© 2012 OSA

OCIS Codes
(110.0180) Imaging systems : Microscopy
(170.0180) Medical optics and biotechnology : Microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: January 4, 2012
Revised Manuscript: February 2, 2012
Manuscript Accepted: February 8, 2012
Published: February 22, 2012

Virtual Issues
Vol. 7, Iss. 4 Virtual Journal for Biomedical Optics

Yasunori Nawa, Wataru Inami, Akito Chiba, Atsushi Ono, Atsuo Miyakawa, Yoshimasa Kawata, Sheng Lin, and Susumu Terakawa, "Dynamic and high-resolution live cell imaging by direct electron beam excitation," Opt. Express 20, 5629-5635 (2012)

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  1. J. Lippincott-Schwartz, E. Snapp, and A. Kenworthy, “Studying protein dynamics in living cells,” Nat. Rev. Mol. Cell Biol. 2(6), 444–456 (2001). [CrossRef] [PubMed]
  2. R. Y. Tsien, “Imagining imaging’s future,” Nat. Rev. Mol. Cell Biol. 5(Suppl), SS16–SS21 (2003). [PubMed]
  3. M. Fernández-Suárez and A. Y. Ting, “Fluorescent probes for super-resolution imaging in living cells,” Nat. Rev. Mol. Cell Biol. 9(12), 929–943 (2008). [CrossRef] [PubMed]
  4. B. Hein, K. I. Willig, C. A. Wurm, V. Westphal, S. Jakobs, and S. W. Hell, “Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins,” Biophys. J. 98(1), 158–163 (2010). [CrossRef] [PubMed]
  5. A. Dani, B. Huang, J. Bergan, C. Dulac, and X. Zhuang, “Superresolution imaging of chemical synapses in the brain,” Neuron 68(5), 843–856 (2010). [CrossRef] [PubMed]
  6. A. Matsuda, L. Shao, J. Boulanger, C. Kervrann, P. M. Carlton, P. Kner, D. Agard, and J. W. Sedat, “Condensed mitotic chromosome structure at nanometer resolution using PALM and EGFP- histones,” PLoS ONE 5(9), e12768 (2010). [CrossRef] [PubMed]
  7. A. Sali, R. Glaeser, T. Earnest, and W. Baumeister, “From words to literature in structural proteomics,” Nature 422(6928), 216–225 (2003). [CrossRef] [PubMed]
  8. W. Baumeister, “A voyage to the inner space of cells,” Protein Sci. 14(1), 257–269 (2005). [CrossRef] [PubMed]
  9. D. Ripper, H. Schwarz, and Y. D. Stierhof, “Cryo-section immunolabelling of difficult to preserve specimens: advantages of cryofixation, freeze-substitution and rehydration,” Biol. Cell 100(2), 109–123 (2008). [CrossRef] [PubMed]
  10. N. de Jonge, N. Poirier-Demers, H. Demers, D. B. Peckys, and D. Drouin, “Nanometer-resolution electron microscopy through micrometers-thick water layers,” Ultramicroscopy 110(9), 1114–1119 (2010). [CrossRef] [PubMed]
  11. O. Cohen, R. Beery, S. Levit, J. Ilany, I. Schwartz, M. Shabtai, D. Anaby, D. Cohen, R. Alfici, A. Czerniak, and A. Karasik, “Scanning electron microscopy of thyroid cells under fully hydrated conditions--a novel technique for a seasoned procedure: a brief observation,” Thyroid 16(10), 997–1001 (2006). [CrossRef] [PubMed]
  12. W. Inami, K. Nakajima, A. Miyakawa, and Y. Kawata, “Electron beam excitation assisted optical microscope with ultra-high resolution,” Opt. Express 18(12), 12897–12902 (2010). [CrossRef] [PubMed]
  13. U. Fano, “A theory on cathode luminescence,” Phys. Rev. 58(6), 544–553 (1940). [CrossRef]
  14. J. Menniger, U. Jahn, O. Brandt, H. Yang, and K. Ploog, “Identification of optical transitions in cubic and hexagonal GaN by spatially resolved cathodoluminescence,” Phys. Rev. B Condens. Matter 53(4), 1881–1885 (1996). [CrossRef] [PubMed]
  15. G. Arnold, ed., Cathodoluminescence and Its Application in the Planetary Sciences, 1st ed. (Springer, 2009).
  16. F. J. G. de Abajo, “Optical excitations in electron microscopy,” Rev. Mod. Phys. 82(1), 209–275 (2010). [CrossRef]
  17. D. C. Joy, Monte Carlo Modeling for Electron Microscopy and Microanalysis (Oxford Univ. Press, 1995).
  18. L. N. Dem’yanets, L. E. Li, and T. G. Uvarova, “Hydrothermal synthesis and cathodoluminescence of ZnO crystalline powders and coatings,” J. Cryst. Growth 287(1), 23–27 (2006). [CrossRef]
  19. K. Senthilkumar, O. Senthilkumar, K. Yamauchi, M. Sato, S. Morito, T. Ohba, M. Nakamura, and Y. Fujita, “Preparation of ZnO nanoparticles for bio-imaging applications,” Phys. Status Solidi B 246(4), 885–888 (2009). [CrossRef]
  20. P. J. Fisher, W. S. Wessels, A. B. Dietz, and F. G. Prendergast, “Enhanced biological cathodoluminescence,” Opt. Commun. 281(7), 1901–1908 (2008). [CrossRef]
  21. E. Kimura, T. Sekiguchi, H. Oikawa, J. Niitsuma, Y. Nakayama, H. Suzuki, M. Kimura, K. Fujii, and T. Ushiki, “Cathodoluminescence imaging for identifying uptaken fluorescence materials in Kupffer cells using scanning electron microscopy,” Arch. Histol. Cytol. 67(3), 263–270 (2004). [CrossRef] [PubMed]
  22. P. V. C. Hough, W. R. McKinney, M. C. Ledbeter, R. E. Pollack, and H. W. Moos, “Identification of biological molecules in situ at high resolution via the fluorescence excited by a scanning electron beam,” Proc. Natl. Acad. Sci. U.S.A. 73(2), 317–321 (1976). [CrossRef] [PubMed]

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