Abstract

Luminescent markers play a key role in imaging techniques for life science since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO3 nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and broad flexibility in the choice of excitation wavelength. We obtained monodispersed BaTiO3 nanoparticles in colloidal suspensions by coating the particle surface with amine groups. We characterized the SHG efficiency of 90-nm BaTiO3 particles experimentally and theoretically. Moreover, we use the coherent SHG signal from BaTiO3 nanoparticles for three-dimensional (3D) imaging without scanning. We built a harmonic holographic (H2) microscope which records digital holograms at the second harmonic frequency. For the first time, high-resolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H2 microscope.

© 2009 Optical Society of America

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2008 (3)

Y. Pu, M. Centurion, and D. Psaltis, "Harmonic holography: a new holographic principle," Appl. Opt. 47, A103-A110 (2008).
[CrossRef] [PubMed]

J. Rosen, and G. Brooker, "Non-scanning motionless fluorescence three-dimensional holographic microscopy," Nat. Photonics 2, 190-195 (2008).
[CrossRef]

A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
[CrossRef]

2007 (4)

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
[CrossRef] [PubMed]

N. Sandeau, L. Le Xuan, D. Chauvat, C. Zhou, J. F. Roch, and S. Brasselet, "Defocused imaging of second harmonic generation from a single nanocrystal," Opt. Express 15, 16051-16060 (2007).
[CrossRef] [PubMed]

P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
[CrossRef]

2006 (5)

M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
[CrossRef] [PubMed]

M. Centurion, Y. Pu, and D. Psaltis, "Holographic capture of femtosecond pulse propagation," J. Appl. Phys. 100, 063104 (2006).
[CrossRef]

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
[CrossRef]

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
[CrossRef]

B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien, "Review - The fluorescent toolbox for assessing protein location and function," Science 312, 217-224 (2006).
[CrossRef] [PubMed]

2005 (1)

2004 (2)

S. Brasselet, V. Le Floc'h, F. Treussart, J. F. Roch, J. Zyss, E. Botzung-Appert, and A. Ibanez, "In situ diagnostics of the crystalline nature of single organic nanocrystals by nonlinear microscopy," Phys. Rev. Lett. 92, 207401 (2004).
[CrossRef] [PubMed]

Y. Pu and H. Meng, "Intrinsic speckle noise in off-axis particle holography," J. Opt. Soc. Am. A-Opt. Image Sci. Vis. 21, 1221-1230 (2004).
[CrossRef] [PubMed]

2003 (3)

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, "Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat Biotech 21, 1356-1360 (2003).
[CrossRef]

A. Miyawaki, A. Sawano, and T. Kogure, "Lighting up cells: labelling proteins with fluorophores," Nature Cell Biol. 5, S1-S7 (2003).

2002 (3)

J. C. Johnson, H. Q. Yan, R. D. Schaller, P. B. Petersen, P. D. Yang, and R. J. Saykally, "Near-field imaging of nonlinear optical mixing in single zinc oxide nanowires," Nano Lett. 2, 279-283 (2002).
[CrossRef]

S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
[CrossRef]

B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
[CrossRef] [PubMed]

2001 (1)

A. A. Heikal, S. T. Hess, and W. W. Webb, "Multiphoton molecular spectroscopy and excited-state dynamics of enhanced green fluorescent protein (EGFP): acid-base specificity," Chem. Phys. 274, 37-55 (2001).
[CrossRef]

2000 (1)

G. H. Patterson and D. W. Piston, "Photobleaching in two-photon excitation microscopy," Biophys. J. 78, 2159-2162 (2000).
[CrossRef] [PubMed]

1999 (1)

P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, "High-resolution nonlinear optical imaging of live cells by second harmonic generation," Biophys. J. 77, 3341-3349 (1999).
[CrossRef] [PubMed]

1998 (4)

W. C. W. Chan, and S. M. Nie, "Quantum dot bioconjugates for ultrasensitive nonisotopic detection," Science 281, 2016-2018 (1998).
[CrossRef] [PubMed]

B. A. Griffin, S. R. Adams, and R. Y. Tsien, "Specific covalent labeling of recombinant protein molecules inside live cells," Science 281, 269-272 (1998).
[CrossRef] [PubMed]

J. Chen, S. Machida, and Y. Yamamoto, "Simultaneous measurement of amplitude and phase in surface second-harmonic generation," Opt. Lett. 23, 676-678 (1998).
[CrossRef]

M. A. Albota, C. Xu, and W. W. Webb, "Two-photon fluorescence excitation cross sections of biomolecular probes from 690 to 960 nm," Appl. Opt. 37, 7352-7356 (1998).
[CrossRef]

1997 (3)

1995 (1)

1994 (1)

M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
[CrossRef] [PubMed]

Adams, S. R.

B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien, "Review - The fluorescent toolbox for assessing protein location and function," Science 312, 217-224 (2006).
[CrossRef] [PubMed]

B. A. Griffin, S. R. Adams, and R. Y. Tsien, "Specific covalent labeling of recombinant protein molecules inside live cells," Science 281, 269-272 (1998).
[CrossRef] [PubMed]

Albota, M. A.

Bonacina, L.

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

Boutou, V.

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

Brasselet, S.

N. Sandeau, L. Le Xuan, D. Chauvat, C. Zhou, J. F. Roch, and S. Brasselet, "Defocused imaging of second harmonic generation from a single nanocrystal," Opt. Express 15, 16051-16060 (2007).
[CrossRef] [PubMed]

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
[CrossRef]

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
[CrossRef]

S. Brasselet, V. Le Floc'h, F. Treussart, J. F. Roch, J. Zyss, E. Botzung-Appert, and A. Ibanez, "In situ diagnostics of the crystalline nature of single organic nanocrystals by nonlinear microscopy," Phys. Rev. Lett. 92, 207401 (2004).
[CrossRef] [PubMed]

Brivanlou, A. H.

B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
[CrossRef] [PubMed]

Brooker, G.

J. Rosen, and G. Brooker, "Non-scanning motionless fluorescence three-dimensional holographic microscopy," Nat. Photonics 2, 190-195 (2008).
[CrossRef]

Bruchez, M. P.

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
[CrossRef] [PubMed]

Budde, F.

S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
[CrossRef]

Campagnola, P. J.

P. J. Campagnola and L. M. Loew, "Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat Biotech 21, 1356-1360 (2003).
[CrossRef]

P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, "High-resolution nonlinear optical imaging of live cells by second harmonic generation," Biophys. J. 77, 3341-3349 (1999).
[CrossRef] [PubMed]

Centurion, M.

Y. Pu, M. Centurion, and D. Psaltis, "Harmonic holography: a new holographic principle," Appl. Opt. 47, A103-A110 (2008).
[CrossRef] [PubMed]

M. Centurion, Y. Pu, and D. Psaltis, "Holographic capture of femtosecond pulse propagation," J. Appl. Phys. 100, 063104 (2006).
[CrossRef]

Chalfie, M.

M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
[CrossRef] [PubMed]

Chan, W. C. W.

W. C. W. Chan, and S. M. Nie, "Quantum dot bioconjugates for ultrasensitive nonisotopic detection," Science 281, 2016-2018 (1998).
[CrossRef] [PubMed]

Chauvat, D.

N. Sandeau, L. Le Xuan, D. Chauvat, C. Zhou, J. F. Roch, and S. Brasselet, "Defocused imaging of second harmonic generation from a single nanocrystal," Opt. Express 15, 16051-16060 (2007).
[CrossRef] [PubMed]

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
[CrossRef]

Chen, J.

Clark, S. W.

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
[CrossRef] [PubMed]

Clement, R.

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
[CrossRef]

Colomb, T.

Courvoisier, F.

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

Cubitt, A. B.

R. M. Dickson, A. B. Cubitt, R. Y. Tsien, and W. E. Moerner, "On/off blinking and switching behaviour of single molecules of green fluorescent protein," Nature 388, 355-358 (1997).
[CrossRef]

Cuche, E.

Delahaye, E.

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
[CrossRef]

Depeursinge, C.

Dickson, R. M.

R. M. Dickson, A. B. Cubitt, R. Y. Tsien, and W. E. Moerner, "On/off blinking and switching behaviour of single molecules of green fluorescent protein," Nature 388, 355-358 (1997).
[CrossRef]

Dohnke, I.

S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
[CrossRef]

Dubertret, B.

B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
[CrossRef] [PubMed]

Ellisman, M. H.

B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien, "Review - The fluorescent toolbox for assessing protein location and function," Science 312, 217-224 (2006).
[CrossRef] [PubMed]

Emery, Y.

Euskirchen, G.

M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
[CrossRef] [PubMed]

Extermann, J.

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

Frank, H.

M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
[CrossRef] [PubMed]

Gacoin, T.

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
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L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
[CrossRef]

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B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien, "Review - The fluorescent toolbox for assessing protein location and function," Science 312, 217-224 (2006).
[CrossRef] [PubMed]

Gratton, E.

Griffin, B. A.

B. A. Griffin, S. R. Adams, and R. Y. Tsien, "Specific covalent labeling of recombinant protein molecules inside live cells," Science 281, 269-272 (1998).
[CrossRef] [PubMed]

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P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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A. A. Heikal, S. T. Hess, and W. W. Webb, "Multiphoton molecular spectroscopy and excited-state dynamics of enhanced green fluorescent protein (EGFP): acid-base specificity," Chem. Phys. 274, 37-55 (2001).
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Hertz, H. M.

Hess, S. T.

A. A. Heikal, S. T. Hess, and W. W. Webb, "Multiphoton molecular spectroscopy and excited-state dynamics of enhanced green fluorescent protein (EGFP): acid-base specificity," Chem. Phys. 274, 37-55 (2001).
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M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
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Johnson, J. C.

J. C. Johnson, H. Q. Yan, R. D. Schaller, P. B. Petersen, P. D. Yang, and R. J. Saykally, "Near-field imaging of nonlinear optical mixing in single zinc oxide nanowires," Nano Lett. 2, 279-283 (2002).
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P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
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Kim, P.

P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
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A. Miyawaki, A. Sawano, and T. Kogure, "Lighting up cells: labelling proteins with fluorophores," Nature Cell Biol. 5, S1-S7 (2003).

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Kuzmin, A. N.

A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
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L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
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M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
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L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
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Ledoux, I.

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
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P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, "High-resolution nonlinear optical imaging of live cells by second harmonic generation," Biophys. J. 77, 3341-3349 (1999).
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B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
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Liphardt, J.

Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
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P. J. Campagnola and L. M. Loew, "Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms," Nat Biotech 21, 1356-1360 (2003).
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M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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Machida, S.

Magistretti, P. J.

Mailander, V.

M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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Mantulin, W. W.

Marder, S. R.

P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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Marquier, F.

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
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Meng, H.

Y. Pu and H. Meng, "Intrinsic speckle noise in off-axis particle holography," J. Opt. Soc. Am. A-Opt. Image Sci. Vis. 21, 1221-1230 (2004).
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A. Miyawaki, A. Sawano, and T. Kogure, "Lighting up cells: labelling proteins with fluorophores," Nature Cell Biol. 5, S1-S7 (2003).

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R. M. Dickson, A. B. Cubitt, R. Y. Tsien, and W. E. Moerner, "On/off blinking and switching behaviour of single molecules of green fluorescent protein," Nature 388, 355-358 (1997).
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Mugnier, Y.

L. Bonacina, Y. Mugnier, F. Courvoisier, R. Le Dantec, J. Extermann, Y. Lambert, V. Boutou, C. Galez, and J. P. Wolf, "Polar Fe(IO3)(3) nanocrystals as local probes for nonlinear microscopy," Appl. Phys.B-Lasers Opt. 87, 399-403 (2007).
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M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
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B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
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B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
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M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
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A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
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Onorato, R. M.

Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
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Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
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L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
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Perry, J. W.

P. Kim, S. C. Jones, P. J. Hotchkiss, J. N. Haddock, B. Kippelen, S. R. Marder, and J. W. Perry, "Phosphonic acid-modified barium titanate polymer nanocomposites with high permittivity and dielectric strength," Adv. Mater. 19, 1001-1005 (2007).
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J. C. Johnson, H. Q. Yan, R. D. Schaller, P. B. Petersen, P. D. Yang, and R. J. Saykally, "Near-field imaging of nonlinear optical mixing in single zinc oxide nanowires," Nano Lett. 2, 279-283 (2002).
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G. H. Patterson and D. W. Piston, "Photobleaching in two-photon excitation microscopy," Biophys. J. 78, 2159-2162 (2000).
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Prasad, P. N.

A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
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M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
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Radenovic, A.

Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
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Rappaz, B.

Rechsteiner, P.

S. Kluge, F. Budde, I. Dohnke, P. Rechsteiner, and J. Hulliger, "Phase-sensitive second-harmonic microscopy reveals polarity of topologically centrosymmetric molecular crystals," Appl. Phys. Lett. 81, 247-249 (2002).
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N. Sandeau, L. Le Xuan, D. Chauvat, C. Zhou, J. F. Roch, and S. Brasselet, "Defocused imaging of second harmonic generation from a single nanocrystal," Opt. Express 15, 16051-16060 (2007).
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J. Rosen, and G. Brooker, "Non-scanning motionless fluorescence three-dimensional holographic microscopy," Nat. Photonics 2, 190-195 (2008).
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A. V. Kachynski, A. N. Kuzmin, M. Nyk, I. Roy, and P. N. Prasad, "Zinc oxide nanocrystals for nonresonant nonlinear optical microscopy in biology and medicine," J. Phys. Chem. C 112, 10721-10724 (2008).
[CrossRef]

Sandeau, N.

Sawano, A.

A. Miyawaki, A. Sawano, and T. Kogure, "Lighting up cells: labelling proteins with fluorophores," Nature Cell Biol. 5, S1-S7 (2003).

Saykally, R. J.

Y. Nakayama, P. J. Pauzauskie, A. Radenovic, R. M. Onorato, R. J. Saykally, J. Liphardt, and P. D. Yang, "Tunable nanowire nonlinear optical probe," Nature 447, 1098-1101 (2007).
[CrossRef] [PubMed]

J. C. Johnson, H. Q. Yan, R. D. Schaller, P. B. Petersen, P. D. Yang, and R. J. Saykally, "Near-field imaging of nonlinear optical mixing in single zinc oxide nanowires," Nano Lett. 2, 279-283 (2002).
[CrossRef]

Schaller, R. D.

J. C. Johnson, H. Q. Yan, R. D. Schaller, P. B. Petersen, P. D. Yang, and R. J. Saykally, "Near-field imaging of nonlinear optical mixing in single zinc oxide nanowires," Nano Lett. 2, 279-283 (2002).
[CrossRef]

Schilling, B. W.

Schrezenmeier, H.

M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
[CrossRef] [PubMed]

Shinoda, K.

Skourides, P.

B. Dubertret, P. Skourides, D. J. Norris, V. Noireaux, A. H. Brivanlou, and A. Libchaber, "In vivo imaging of quantum dots encapsulated in phospholipid micelles," Science 298, 1759-1762 (2002).
[CrossRef] [PubMed]

So, P. T. C.

Storrie, B.

Suzuki, Y.

Tancrez, N.

E. Delahaye, N. Tancrez, T. Yi, I. Ledoux, J. Zyss, S. Brasselet, and R. Clement, "Second harmonic generation from individual hybrid MnPS3-based nanoparticles investigated by nonlinear microscopy," Chem. Phys. Lett. 429, 533-537 (2006).
[CrossRef]

Tard, C.

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
[CrossRef]

Treussart, F.

L. L. Xuan, S. Brasselet, F. Treussart, J. F. Roch, F. Marquier, D. Chauvat, S. Perruchas, C. Tard, and T. Gacoin, "Balanced homodyne detection of second-harmonic generation from isolated subwavelength emitters," Appl. Phys. Lett. 89, 121118 (2006).
[CrossRef]

Tsien, R. Y.

B. N. G. Giepmans, S. R. Adams, M. H. Ellisman, and R. Y. Tsien, "Review - The fluorescent toolbox for assessing protein location and function," Science 312, 217-224 (2006).
[CrossRef] [PubMed]

B. A. Griffin, S. R. Adams, and R. Y. Tsien, "Specific covalent labeling of recombinant protein molecules inside live cells," Science 281, 269-272 (1998).
[CrossRef] [PubMed]

R. M. Dickson, A. B. Cubitt, R. Y. Tsien, and W. E. Moerner, "On/off blinking and switching behaviour of single molecules of green fluorescent protein," Nature 388, 355-358 (1997).
[CrossRef]

Tu, Y.

M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
[CrossRef] [PubMed]

Walther, P.

M. R. Lorenz, V. Holzapfel, A. Musyanovych, K. Nothelfer, P. Walther, H. Frank, K. Landfester, H. Schrezenmeier, and V. Mailander, "Uptake of functionalized, fluorescent-labeled polymeric particles in different cell lines and stem cells," Biomaterials 27, 2820-2828 (2006).
[CrossRef] [PubMed]

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M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, and D. C. Prasher, "Green fluorescent protein as a marker for gene-expression," Science 263, 802-805 (1994).
[CrossRef] [PubMed]

Webb, W. W.

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
[CrossRef] [PubMed]

A. A. Heikal, S. T. Hess, and W. W. Webb, "Multiphoton molecular spectroscopy and excited-state dynamics of enhanced green fluorescent protein (EGFP): acid-base specificity," Chem. Phys. 274, 37-55 (2001).
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Wei, M. D.

P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, "High-resolution nonlinear optical imaging of live cells by second harmonic generation," Biophys. J. 77, 3341-3349 (1999).
[CrossRef] [PubMed]

Williams, R. M.

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
[CrossRef] [PubMed]

Wise, F. W.

D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, "Water-soluble quantum dots for multiphoton fluorescence imaging in vivo," Science 300, 1434-1436 (2003).
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Figures (8)

Fig. 1.
Fig. 1.

Dynamic light scattering measurement of 90-nm BaTiO3 particles colloidal suspension. Inset: Scanning electron microscope image of a single 90-nm BaTiO3 particle.

Fig. 2.
Fig. 2.

Power dependence of the SHG signal from BaTiO3 nanoparticles in double logarithmic scale. The squares are measured results and the solid line is the linear fit with the slope of 2.2. The inset shows the SHG optical spectrum centered at 400.6 nm with a full width half maximum of 5 nm.

Fig. 3.
Fig. 3.

Polarization dependent SHG response measured from an isolated BaTiO3 nanocrystal. Dots: experimental data. Line: Theoretical calculation based on the second order susceptibility of a bulk BaTiO3 crystal oriented at ϕ0 = 90 degree and θ0 = 30, 50, 70, 90 degree respectively. The red arrow in the polar diagram indicates the projection of the c-axis of the nanocrystal on the XY plane.

Fig. 4.
Fig. 4.

H2 microscope experimental setup. BS1 and BS2, beam splitters; M, mirror; L1 and L2, lens; S, sample; OBJ, microscope objective; BE, beam expander; F, band-pass filter centered at 400 nm. BS1 splits the laser into signal and reference beams. In the signal arm, L1 slightly focuses the excitation beam into the sample with SHRIMPs. OBJ and L2 form a 4F imaging system to collect and optically magnify the SHG image of SHRIMP. The EMCCD is placed away from the 4F imaging plane. A band-pass filter is placed in front of the EMCCD to remove the excitation from the SHG signal. The reference beam goes through a translation stage and a BBO crystal so that the coherent reference SHG laser pulses are generated and can be temporally and spatially overlapped with the signal on the EMCCD. The signal and reference beams are combined collinearly by BS2 and therefore an on-axis digital hologram is recorded on the EMCCD.

Fig. 5.
Fig. 5.

Spatial resolution of the H2 microscope measured by imaging an isolated BaTiO3 particle. (a) SHG intensity image measured by a conventional microscope. (b) Holographic reconstructed intensity image at the object plane. (c) Cross section of a stack of holographic reconstructed intensity images along the axial direction. The scale bars are 2 μm.

Fig. 6.
Fig. 6.

Images of SHRIMPs embedded in PDMS. (a)–(d): SHG images of SHRIMPs on four different planes, focusing by moving the optics with a conventional microscope. (e)–(h): Holographic reconstructed images of SHRIMPs on four corresponding planes, focusing by digital reconstruction. The relative depths of these four planes are 0, 9.4, 17.2, and 20.3 μm respectively. Scale bars are 2 μm.

Fig. 7.
Fig. 7.

Confocal section images of a calcein-stained HeLa cell (in red) with SHRIMPs (in green) inside it. The excitation consisted of femtosecond laser pulses centered at 800-nm wavelength. Calcein emits two-photon fluorescence signal (in red) while SHRIMPs emit SHG signal (in green). It is clear that the SHRIMPs entered the cell through endocytosis. The scale bar is 5μm.

Fig. 8.
Fig. 8.

Images of SHRIMPs non-specifically labeling HeLa cells (a) Superposition of the bright field transmission image of a HeLa cell (in red) and the SHG image of SHRIMPs (in green) taken by a conventional microscope. Six SHRIMPs clusters assigned with numbers were under analysis. Some of them cannot be seen clearly because they are out of focus. (b)–(d) H2 reconstructed images at three different planes with relative depths of 0 μm, 3.12 μm and 6.24 μm respectively. The white arrows show the SHRIMPs that are on focus, while the gray arrows show the SHRIMPs that are out of focus. The SHRIMP labeled with number 4 is a big cluster so that it is bright both in (b) and (c). (e) Normalized intensity line profiles of the six clusters when they are on focus through digital reconstruction. The scale bars are 5 μm.

Equations (4)

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W 2 P = σ 2 P I 1 P 2 .
P ( ω , 2 ω , ) = χ ( 1 ) · E 1 ( ω ) + χ ( 2 ) · E 1 ( ω ) · E 1 ( ω ) + .
P ( 2 ω ) = χ ( 2 ) · E 1 ( ω ) · E 1 ( ω ) .
W 0 = c k 4 12 π ε 0 [ d i ( 2 ω ) ] 2 ,

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