Abstract

An environmental cell with a 50-nm-thick cathodolumi-nescent window was attached to a scanning electron microscope, and diffraction-unlimited near-field optical imaging of unstained living human lung epithelial cells in liquid was demonstrated. Electrons with energies as low as 0.8 – 1.2 kV are sufficiently blocked by the window without damaging the specimens, and form a sub-wavelength-sized illumination light source. A super-resolved optical image of the specimen adhered to the opposite window surface was acquired by a photomultiplier tube placed below. The cells after the observation were proved to stay alive. The image was formed by enhanced dipole radiation or energy transfer, and features as small as 62 nm were resolved.

© 2013 Optical Society of America

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2013

Y. Cotte, F. Toy, P. Jourdain, N. Pavillon, D. Boss, P. Magistretti, P. Marquet, and C. Depeursinge, “Marker-free phase nanoscopy,” Nat. Photonics7, 113–117 (2013).
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[CrossRef]

A. Chiba, S. Tanaka, W. Inami, A. Sugita, K. Takada, and Y. Kawata, “Amorphous silicon nitride thin films implanted with cerium ions for cathodoluminescent light source,” Opt. Mater.35, 1887–1889 (2013).
[CrossRef]

2012

2011

N. Hanagata, F. Zhuang, S. Connolly, J. Li, N. Ogawa, and M. Xu, “Molecular responses of human lung epithelial cells to the toxicity of copper oxide nanoparticles inferred from whole genome expression analysis,”ACS Nano5, 9326–9338 (2011).
[CrossRef] [PubMed]

H. Niioka, T. Furukawa, M. Ichimiya, M. Ashida, T. Araki, and M. Hashimoto, “Multicolor cathodoluminescence microscopy for biological imaging with nanophosphors,” Appl. Phys. Express4, 112402 (2011).
[CrossRef]

N. de Jonge and F. M. Ross, “Electron miocroscopy of specimens in liquid,” Nat. Nanotechnol.6, 695–704 (2011).
[CrossRef] [PubMed]

D. B. Peckys, P. Mazur, K. L. Gould, and N. de Jonge, “Fully hydrated yeast cells imaged with electron microscopy,” Biophys. J.100, 2522–2529 (2011).
[CrossRef] [PubMed]

2010

H. Nishiyama, M. Suga, T. Ogura, Y. Maruyama, M. Koizumi, K. Mio, S. Kitamura, and C. Sato, “Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film,” J. Struct. Biol.169, 438–449 (2010).
[CrossRef] [PubMed]

T. Oto, R. G. Banal, K. Kataoka, M. Funato, and Y. Kawakami, “100 mW deep-ultraviolet emission from aluminium-nitride-based quantum wells pumped by an electron beam,”Nat. Photonics4, 767–771 (2010).
[CrossRef]

M. Xu, D. Fujita, S. Kajiwara, T. Minowa, X. Li, T. Takemura, H. Iwai, and N. Hanagata, “Contribution of physicochemical characteristics of nano-oxides to cytotoxicity,” Biomaterials31, 8022–8031 (2010).
[CrossRef] [PubMed]

W. Inami, K. Nakajima, A. Miyakawa, and Y. Kawata, “Electron beam excitation assisted optical microscope with ultra-high resolution,” Opt. Express18, 12897–12902 (2010).
[CrossRef] [PubMed]

2009

J. Han, J. An, R. N. Jana, K. Jung, J. Do, S. Pyo, and C. Im, “Charge carrier photogeneration and hole transport properties of blends of a π-conjugated polymer and an organic-inorganic hybrid material,” Macromol. Res.17, 894–900 (2009).
[CrossRef]

T. Ogura, “Measurement of the unstained biological sample by a novel scanning electron generation X-ray microscope based on SEM,” Biochem. Biophys. Res. Commun.385, 624–629 (2009).
[CrossRef] [PubMed]

R. R. Lunt, N. C. Giebink, A. A. Belak, J. B. Benziger, and S. R. Forrest, “Exciton diffusion lengths of organic semiconductor thin films measured by spectrally resolved photoluminescence quenching,” J. Appl. Phys.105, 053711 (2009).
[CrossRef]

B. Barwick, D. J. Flannigan, and A. H. Zewail, “Photon-induced near-field electron microscopy,” Nature462, 902–906 (2009).
[CrossRef] [PubMed]

N. de Jonge, D. B. Peckys, G. J. Kremers, and D. W. Piston, “Electron microscopy of whole cells in liquid with nanometer resolution,” Proc. Natl. Acad. Sci. U. S. A.106, 2159–2164 (2009).
[CrossRef] [PubMed]

R. Böhme, M. Richter, D. Cialla, P. Rösch, V. Deckert, and J. Popp, “Towards a specific characterisation of components on a cell surface – combined TERS – investigations of lipids and human cells,” J. Raman Spectrosc.40, 1452–1457 (2009).
[CrossRef]

2008

C. Höppener and L. Novotny, “Imaging of membrane proteins using antenna-based optical microscopy,” Nanotechnology19, 384012 (2008).
[CrossRef] [PubMed]

K.-L. Liu, C.-C. Wu, Y.-J. Huang, H.-L. Peng, W.-Y. Chang, P. Chang, L. Hsu, and T.-R. Yew, “Novel microchip for in situ TEM imaging of living organisms and bio-reactions in aqueous conditions,” Lab Chip8, 1915–1921 (2008).
[CrossRef] [PubMed]

2007

S. W. Hell, “Far-field optical nanoscopy,” Science316, 1153–1158 (2007).
[CrossRef] [PubMed]

J. Nelayah, M. Kociak, O. Stéphan, F. J. García de Abajo, M. Tencé, L. Henrard, D. Taverna, I. Pastoriza-Santos, L. M. Liz-Marzán, and C. Colliex, “Mapping surface plasmons on a single metallic nanoparticle,” Nat. Phys.3, 348–353 (2007).
[CrossRef]

D. Drouin, A. R. Couture, D. Joly, X. Tastet, V. Aimez, and R. Gauvin, “CASINO V2.42—a fast and easy-to-use modeling tool for scanning electron microscopy and microanalysis users,” Scanning29, 92–101 (2007).
[CrossRef] [PubMed]

H. Mertens, A. F. Koenderink, and A. Polman, “Plasmon-enhanced luminescence near noble-metal nanospheres: comparison of exact theory and an improved Gersten and Nitzan model,” Phys. Rev. B76, 115123 (2007).
[CrossRef]

2006

E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent protains at nanometer resolution,” Science313, 1642–1645 (2006).
[CrossRef] [PubMed]

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3, 793–795 (2006).
[CrossRef] [PubMed]

2005

M. G. L. Gustafsson, “Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution,” Proc. Natl. Acad. Sci. U. S. A.102, 13081–13086 (2005).
[CrossRef] [PubMed]

J. Niitsuma, H. Oikawa, E. Kimura, T. Ushiki, and T. Sekiguchi, “Cathodoluminescence investigation of organic materials,” J. Electron Microsc.54, 325–330 (2005).
[CrossRef]

A. Kubo, K. Onda, H. Petek, Z. Sun, Y. S. Jung, and H. K. Kim, “Femtosecond imaging of surface plasmon dynamics in a nanostructured silver film,” Nano Lett.5, 1123–1127 (2005).
[CrossRef] [PubMed]

2004

S. Thiberge, A. Nechushtan, D. Sprinzak, O. Gileadi, V. Behar, O. Zik, Y. Chowers, S. Michaeli, J. Schlessinger, and E. Moses, “Scanning electron microscopy of cells and tissues under fully hydrated conditions,” Proc. Natl. Acad. Sci. U. S. A.101, 3346–3351 (2004).
[CrossRef] [PubMed]

M. Koopman, A. Cambi, B. I. de Bakker, B. Joosten, C. G. Figdor, N. F. van Hulst, and M. F. Garcia-Parajo, “Near-field scanning optical microscopy in liquid for high resolution single molecule detection on dendritic cells,” FEBS Lett.573, 6–10 (2004).
[CrossRef] [PubMed]

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, 263–270 (2004).
[CrossRef] [PubMed]

K. Nakaoka, J. Ueyama, and K. Ogura, “Photoelectrochemical behavior of electrodeposited CuO and Cu2O thin films on conducting substrates,” J. Electrochem. Soc.151, C661–C665 (2004).
[CrossRef]

2001

N. Yamamoto, K. Araya, and F. J. García de Abajo, “Photon emission from silver particles induced by a high-energy electron beam,” Phys. Rev. B64, 205419 (2001).
[CrossRef]

2000

N. Hayazawa, Y. Inouye, Z. Sekkat, and S. Kawata, “Metallized tip amplification of near-field Raman scattering,” Opt. Commun.183, 333–336 (2000).
[CrossRef]

T. A. Klar, S. Jakobs, M. Dyba, A. Egner, and S. W. Hell, “Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission,” Proc. Natl. Acad. Sci. U. S. A.97, 8206–8210 (2000).
[CrossRef] [PubMed]

K. Postava, H. Sueki, M. Aoyama, T. Yamaguchi, Ch. Ino, Y. Igasaki, and M. Horie, “Spectroscopic ellipsometry of epitaxial ZnO layer on sapphire substrate,” J. Appl. Phys.87, 7820–7824 (2000).
[CrossRef]

1998

T. Saiki, K. Nishi, and M. Ohtsu, “Low temperature near-field photoluminescence spectroscopy of InGaAs single quantum dots,”Jpn. J. Appl. Phys.37, 1638–1642 (1998).
[CrossRef]

1996

A. Stoffel, A. Kovács, W. Kronast, and B. Müller, “LPCVD against PECVD for micromechanical applications,” J. Micromech. Microeng.6, 1–13 (1996).
[CrossRef]

1995

T. Funatsu, Y. Harada, M. Tokunaga, K. Saito, and T. Yanagida, “Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution,” Nature374, 555–559 (1995).
[CrossRef] [PubMed]

J. Kido, H. Shionoya, and K. Nagai, “Single-layer white light-emitting organic electroluminescent devices based on dye-dispersed poly(N-vinylcarbazole),” Appl. Phys. Lett.67, 2281–2283 (1995).
[CrossRef]

1984

D. W. Pohl, W. Denk, and M. Lanz, “Optical stethoscopy: image recording with resolution λ/20,” Appl. Phys. Lett.44, 651–653 (1984).
[CrossRef]

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, “Development of a 500 Å spatial resolution light microscope,” Ultramicroscopy13, 227–232 (1984).
[CrossRef]

1978

R. Herbst and D. Hoder, “Cathodoluminescence in biological studies,” Scanning1, 35–41 (1978).
[CrossRef]

V. E. Cosslett, “Radiation damage in the high resolution electron microscopy of biological materials: a review,” J. Microsc.113, 113–129 (1978).
[CrossRef] [PubMed]

1974

D. F. Parsons, V. R. Matricardi, R. C. Moretz, and J. N. Turner, “Electron microscopy and diffraction of wet unstained and unfixed biological objects,” Adv. Biol. Med. Phys.15, 161–270 (1974).
[PubMed]

1973

1972

K. Kanaya and S. Okayama, “Penetration and energy-loss theory of electrons in solid targets,” J. Phys. D Appl. Phys.5, 43–58 (1972).
[CrossRef]

J. E. Mazurkiewicz and P. K. Nakane, “Light and electron microscopic localization of antigens in tissues embedded in polyethylene glycol with a peroxidase-labeled antibody method,” J. Histochem. Cytochem.20, 969–974 (1972).
[CrossRef] [PubMed]

Aimez, V.

D. Drouin, A. R. Couture, D. Joly, X. Tastet, V. Aimez, and R. Gauvin, “CASINO V2.42—a fast and easy-to-use modeling tool for scanning electron microscopy and microanalysis users,” Scanning29, 92–101 (2007).
[CrossRef] [PubMed]

An, J.

J. Han, J. An, R. N. Jana, K. Jung, J. Do, S. Pyo, and C. Im, “Charge carrier photogeneration and hole transport properties of blends of a π-conjugated polymer and an organic-inorganic hybrid material,” Macromol. Res.17, 894–900 (2009).
[CrossRef]

Aoyama, M.

K. Postava, H. Sueki, M. Aoyama, T. Yamaguchi, Ch. Ino, Y. Igasaki, and M. Horie, “Spectroscopic ellipsometry of epitaxial ZnO layer on sapphire substrate,” J. Appl. Phys.87, 7820–7824 (2000).
[CrossRef]

Araki, T.

H. Niioka, T. Furukawa, M. Ichimiya, M. Ashida, T. Araki, and M. Hashimoto, “Multicolor cathodoluminescence microscopy for biological imaging with nanophosphors,” Appl. Phys. Express4, 112402 (2011).
[CrossRef]

Araya, K.

N. Yamamoto, K. Araya, and F. J. García de Abajo, “Photon emission from silver particles induced by a high-energy electron beam,” Phys. Rev. B64, 205419 (2001).
[CrossRef]

Ashida, M.

H. Niioka, T. Furukawa, M. Ichimiya, M. Ashida, T. Araki, and M. Hashimoto, “Multicolor cathodoluminescence microscopy for biological imaging with nanophosphors,” Appl. Phys. Express4, 112402 (2011).
[CrossRef]

Banal, R. G.

T. Oto, R. G. Banal, K. Kataoka, M. Funato, and Y. Kawakami, “100 mW deep-ultraviolet emission from aluminium-nitride-based quantum wells pumped by an electron beam,”Nat. Photonics4, 767–771 (2010).
[CrossRef]

Barwick, B.

B. Barwick, D. J. Flannigan, and A. H. Zewail, “Photon-induced near-field electron microscopy,” Nature462, 902–906 (2009).
[CrossRef] [PubMed]

Bass, M.

M. Bass, Handbook of Optics II, 2 (McGraw-Hill, 1995).

Bates, M.

M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods3, 793–795 (2006).
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P. Wang, M. N. Slipchenko, J. Mitchell, C. Yang, E. O. Potma, X. Xu, and J. X. Cheng, “Far-field imaging of non-fluorescent species with subdiffraction resolution,” Nat. Photonics7, 449–453 (2013).
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T. Funatsu, Y. Harada, M. Tokunaga, K. Saito, and T. Yanagida, “Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution,” Nature374, 555–559 (1995).
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P. Wang, M. N. Slipchenko, J. Mitchell, C. Yang, E. O. Potma, X. Xu, and J. X. Cheng, “Far-field imaging of non-fluorescent species with subdiffraction resolution,” Nat. Photonics7, 449–453 (2013).
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K.-L. Liu, C.-C. Wu, Y.-J. Huang, H.-L. Peng, W.-Y. Chang, P. Chang, L. Hsu, and T.-R. Yew, “Novel microchip for in situ TEM imaging of living organisms and bio-reactions in aqueous conditions,” Lab Chip8, 1915–1921 (2008).
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B. Barwick, D. J. Flannigan, and A. H. Zewail, “Photon-induced near-field electron microscopy,” Nature462, 902–906 (2009).
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N. Hanagata, F. Zhuang, S. Connolly, J. Li, N. Ogawa, and M. Xu, “Molecular responses of human lung epithelial cells to the toxicity of copper oxide nanoparticles inferred from whole genome expression analysis,”ACS Nano5, 9326–9338 (2011).
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S. Thiberge, A. Nechushtan, D. Sprinzak, O. Gileadi, V. Behar, O. Zik, Y. Chowers, S. Michaeli, J. Schlessinger, and E. Moses, “Scanning electron microscopy of cells and tissues under fully hydrated conditions,” Proc. Natl. Acad. Sci. U. S. A.101, 3346–3351 (2004).
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ACS Nano

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M. Xu, D. Fujita, S. Kajiwara, T. Minowa, X. Li, T. Takemura, H. Iwai, and N. Hanagata, “Contribution of physicochemical characteristics of nano-oxides to cytotoxicity,” Biomaterials31, 8022–8031 (2010).
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P. Wang, M. N. Slipchenko, J. Mitchell, C. Yang, E. O. Potma, X. Xu, and J. X. Cheng, “Far-field imaging of non-fluorescent species with subdiffraction resolution,” Nat. Photonics7, 449–453 (2013).
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Nature

B. Barwick, D. J. Flannigan, and A. H. Zewail, “Photon-induced near-field electron microscopy,” Nature462, 902–906 (2009).
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Opt. Express

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