Abstract

Using a total internal reflection fluorescence microscopy (TIRFM) technique to image live cells on a biosurface not only provides an enhanced understanding of cellular functions, but also improves the signal-to-noise ratio of the images. However, the intensity of the fluorescence signal must be increased if a more dynamic biomolecular imaging capability is required. Accordingly, this study presents a surface plasmon-enhanced TIRFM technique in which the fluorescence signals are enhanced via surface plasmons offered by a silver nanolayer. The developed microscopy technique is successfully applied to the real-time observation of the thrombomodulin proteins of live cell membranes. The experimental results and the simulation results demonstrate that the live cell membrane images obtained in the proposed surface plasmon-enhanced TIRFM technique are brighter by approximately one order of magnitude than those provided by conventional TIRFM.

© 2006 Optical Society of America

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  30. M. C. Boffa, B. Burke, and C. C. Haudenschild, "Preservation of thrombomodulin antigen on vascular and extravascular surfaces," J. Histochem. Cytochem. 35, 1267-1276 (1987).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [PubMed]
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    [CrossRef]
  35. S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
    [CrossRef] [PubMed]

2006

P. Anger, P. Bharadwaj, and L. Novotny, "Enhancement and quenching of single-molecule fluorescence," Phys. Rev. Lett. 96, 113002 (2006).
[CrossRef] [PubMed]

2005

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

J. Enderlein and T. Ruckstuhl, "The efficiency of surface-plasmon coupled emission for sensitive fluorescence detection," Opt. Express 13, 8855-8865 (2005).
[CrossRef] [PubMed]

I. H. El-Sayed, X. Huang, and M. A. El-Sayed, "Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer," Nano Lett. 5, 829-835 (2005).
[CrossRef] [PubMed]

S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
[CrossRef] [PubMed]

H. Schneckenburger, "Total internal reflection fluorescence microscopy: technical innovations and novel applications," Curr. Opin. Biotechnol. 16, 13-18 (2005).
[CrossRef] [PubMed]

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

O. Stranik, H. M. McEvoy, C. McDonagh, and B. D. MacCraith, "Plasmonic enhancement of fluorescence for sensor applications," Sens. Actuators B 107, 148-153 (2005).
[CrossRef]

2004

K. Tawa and W. Knoll, "Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy," Nucleic Acids Res. 32, 2372-2377 (2004).
[CrossRef] [PubMed]

F. Yu, B. Persson, S. Lofas, and W. Knoll, "Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level," Anal. Chem. 76, 6765-6770 (2004).
[CrossRef] [PubMed]

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
[CrossRef] [PubMed]

S.-J. Chen, F. C. Chien, G. Y. Lin, and K. C. Lee, "Enhanced the resolution of surface plasmon resonance biosensors by controlling size and distribution of nanoparticles," Opt. Lett. 29, 1390-1392 (2004).
[CrossRef] [PubMed]

2003

H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, "Thrombomodulin-mediated cell adhesion," J. Biol. Chem. 278, 46750-46759 (2003).
[CrossRef] [PubMed]

G. Raschke, S. Kowarik, T. Franzl, C. So1nnichsen, T. A. Klar, J. Feldmann, A. Nichtl, and K. Ku1rzinger, "Biomolecular recognition based on single gold nanoparticle light scattering," Nano Lett. 3, 935-938 (2003).
[CrossRef]

2002

C. D. Geddes and J. R. Lakowicz, "Metal-enhanced fluorescence," J. Fluor. 12, 121-129 (2002).
[CrossRef]

2001

J. R. Lakowicz, "Radiative decay engineering: biophysical and biomedical applications," Anal. Biochem. 298, 1-24 (2001).
[CrossRef] [PubMed]

D. Toomre and D. J. Manstein, "Lighting up the cell surface with evanescent wave microscopy," Trends Cell Biol. 11, 298-303 (2001).
[CrossRef] [PubMed]

D. Axelrod, "Total internal reflection fluorescence microscopy in cell biology," Traffic 2, 764-774 (2001).
[CrossRef] [PubMed]

J. A. Steyer and W. Almers, "A real-time view of within life within 100 nm of the plasma membrane," Nat. Rev. Mol. Cell Biol. 2, 268-276 (2001).
[CrossRef] [PubMed]

2000

S. E. Sund and D. Axelrod, "Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching," Biophys. J. 79, 1655-1669 (2000).
[CrossRef] [PubMed]

T. Liebermann and W. Knoll, "Surface-plasmon field-enhanced fluorescence spectroscopy," Colloids Surf. A 171, 115-130 (2000).
[CrossRef]

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

1999

J. Homola, S. S. Yee, and G. Gauglitz, "Surface plasmon resonance sensors: review," Sens. Actuators B 54, 3-15 (1999).
[CrossRef]

1998

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

1996

W. J. Betz, F. Mao, and C. B. Smith, "Imaging exocytosis and endocytosis," Curr. Opin. Neurobiol. 6, 365-371 (1996).
[CrossRef] [PubMed]

1995

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

1993

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
[CrossRef]

R. M. Fulbright and D. Axelrod, "Dynamics of nonspecific adsorption of insulin to erythrocyte membranes," J. Fluor. 3, 1-16 (1993).
[CrossRef]

1992

G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, "Total internal reflection fluorescence microscopy (TIRFM) II. Topographical mapping of relative cell/substratum separation distances," J. Cell Sci. 103, 491-499 (1992).
[PubMed]

1987

M. C. Boffa, B. Burke, and C. C. Haudenschild, "Preservation of thrombomodulin antigen on vascular and extravascular surfaces," J. Histochem. Cytochem. 35, 1267-1276 (1987).
[CrossRef] [PubMed]

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

1979

Almers, W.

J. A. Steyer and W. Almers, "A real-time view of within life within 100 nm of the plasma membrane," Nat. Rev. Mol. Cell Biol. 2, 268-276 (2001).
[CrossRef] [PubMed]

Anger, P.

P. Anger, P. Bharadwaj, and L. Novotny, "Enhancement and quenching of single-molecule fluorescence," Phys. Rev. Lett. 96, 113002 (2006).
[CrossRef] [PubMed]

Aslan, K.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

Aussenegg, F.

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
[CrossRef]

Axelrod, D.

D. Axelrod, "Total internal reflection fluorescence microscopy in cell biology," Traffic 2, 764-774 (2001).
[CrossRef] [PubMed]

S. E. Sund and D. Axelrod, "Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching," Biophys. J. 79, 1655-1669 (2000).
[CrossRef] [PubMed]

R. M. Fulbright and D. Axelrod, "Dynamics of nonspecific adsorption of insulin to erythrocyte membranes," J. Fluor. 3, 1-16 (1993).
[CrossRef]

Betz, W. J.

W. J. Betz, F. Mao, and C. B. Smith, "Imaging exocytosis and endocytosis," Curr. Opin. Neurobiol. 6, 365-371 (1996).
[CrossRef] [PubMed]

Bharadwaj, P.

P. Anger, P. Bharadwaj, and L. Novotny, "Enhancement and quenching of single-molecule fluorescence," Phys. Rev. Lett. 96, 113002 (2006).
[CrossRef] [PubMed]

Bocchio, N.

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

Boffa, M. C.

M. C. Boffa, B. Burke, and C. C. Haudenschild, "Preservation of thrombomodulin antigen on vascular and extravascular surfaces," J. Histochem. Cytochem. 35, 1267-1276 (1987).
[CrossRef] [PubMed]

Böhrer, H.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Brunner, H.

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
[CrossRef]

Burke, B.

M. C. Boffa, B. Burke, and C. C. Haudenschild, "Preservation of thrombomodulin antigen on vascular and extravascular surfaces," J. Histochem. Cytochem. 35, 1267-1276 (1987).
[CrossRef] [PubMed]

Burmeister, J. S.

G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, "Total internal reflection fluorescence microscopy (TIRFM) II. Topographical mapping of relative cell/substratum separation distances," J. Cell Sci. 103, 491-499 (1992).
[PubMed]

Chen, J.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Chen, S.-J.

Chien, F. C.

Deng, Y.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Deyashiki, Y.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Eagen, C. F.

Ekgasit, S.

S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
[CrossRef] [PubMed]

El-Sayed, I. H.

I. H. El-Sayed, X. Huang, and M. A. El-Sayed, "Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer," Nano Lett. 5, 829-835 (2005).
[CrossRef] [PubMed]

El-Sayed, M. A.

I. H. El-Sayed, X. Huang, and M. A. El-Sayed, "Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer," Nano Lett. 5, 829-835 (2005).
[CrossRef] [PubMed]

Emmert, H.

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Enderlein, J.

Franzl, T.

G. Raschke, S. Kowarik, T. Franzl, C. So1nnichsen, T. A. Klar, J. Feldmann, A. Nichtl, and K. Ku1rzinger, "Biomolecular recognition based on single gold nanoparticle light scattering," Nano Lett. 3, 935-938 (2003).
[CrossRef]

Fulbright, R. M.

R. M. Fulbright and D. Axelrod, "Dynamics of nonspecific adsorption of insulin to erythrocyte membranes," J. Fluor. 3, 1-16 (1993).
[CrossRef]

Gauglitz, G.

J. Homola, S. S. Yee, and G. Gauglitz, "Surface plasmon resonance sensors: review," Sens. Actuators B 54, 3-15 (1999).
[CrossRef]

Geddes, C. D.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

C. D. Geddes and J. R. Lakowicz, "Metal-enhanced fluorescence," J. Fluor. 12, 121-129 (2002).
[CrossRef]

Graeff, H.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Grapa, E.

G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, "Total internal reflection fluorescence microscopy (TIRFM) II. Topographical mapping of relative cell/substratum separation distances," J. Cell Sci. 103, 491-499 (1992).
[PubMed]

Gryczynski, I.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
[CrossRef] [PubMed]

Gryczynski, Z.

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
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Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
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J. Homola, S. S. Yee, and G. Gauglitz, "Surface plasmon resonance sensors: review," Sens. Actuators B 54, 3-15 (1999).
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K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

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K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Huang, H. C.

H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, "Thrombomodulin-mediated cell adhesion," J. Biol. Chem. 278, 46750-46759 (2003).
[CrossRef] [PubMed]

Huang, X.

I. H. El-Sayed, X. Huang, and M. A. El-Sayed, "Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer," Nano Lett. 5, 829-835 (2005).
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H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, "Thrombomodulin-mediated cell adhesion," J. Biol. Chem. 278, 46750-46759 (2003).
[CrossRef] [PubMed]

Kawahara, S.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Klevesath, M.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
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Knobloch, H.

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
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Knoll, W.

S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
[CrossRef] [PubMed]

K. Tawa and W. Knoll, "Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy," Nucleic Acids Res. 32, 2372-2377 (2004).
[CrossRef] [PubMed]

F. Yu, B. Persson, S. Lofas, and W. Knoll, "Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level," Anal. Chem. 76, 6765-6770 (2004).
[CrossRef] [PubMed]

T. Liebermann and W. Knoll, "Surface-plasmon field-enhanced fluorescence spectroscopy," Colloids Surf. A 171, 115-130 (2000).
[CrossRef]

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
[CrossRef]

Kowarik, S.

G. Raschke, S. Kowarik, T. Franzl, C. So1nnichsen, T. A. Klar, J. Feldmann, A. Nichtl, and K. Ku1rzinger, "Biomolecular recognition based on single gold nanoparticle light scattering," Nano Lett. 3, 935-938 (2003).
[CrossRef]

Kreiter, M.

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

Kusano, C.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Kusumoto, H.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Lakowicz, J. R.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
[CrossRef] [PubMed]

C. D. Geddes and J. R. Lakowicz, "Metal-enhanced fluorescence," J. Fluor. 12, 121-129 (2002).
[CrossRef]

J. R. Lakowicz, "Radiative decay engineering: biophysical and biomedical applications," Anal. Biochem. 298, 1-24 (2001).
[CrossRef] [PubMed]

Lee, K. C.

Leitner, A.

H. Knobloch, H. Brunner, A. Leitner, F. Aussenegg, and W. Knoll, "Probing the evanescent field of propagating plasmon surface polaritons by fluorescence and Raman spectroscopies," J. Chem. Phys. 98, 10093-10095 (1993).
[CrossRef]

Liebermann, T.

T. Liebermann and W. Knoll, "Surface-plasmon field-enhanced fluorescence spectroscopy," Colloids Surf. A 171, 115-130 (2000).
[CrossRef]

Lin, G. Y.

Lofas, S.

F. Yu, B. Persson, S. Lofas, and W. Knoll, "Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level," Anal. Chem. 76, 6765-6770 (2004).
[CrossRef] [PubMed]

MacCraith, B. D.

O. Stranik, H. M. McEvoy, C. McDonagh, and B. D. MacCraith, "Plasmonic enhancement of fluorescence for sensor applications," Sens. Actuators B 107, 148-153 (2005).
[CrossRef]

Malicka, J.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
[CrossRef] [PubMed]

Manstein, D. J.

D. Toomre and D. J. Manstein, "Lighting up the cell surface with evanescent wave microscopy," Trends Cell Biol. 11, 298-303 (2001).
[CrossRef] [PubMed]

Mao, F.

W. J. Betz, F. Mao, and C. B. Smith, "Imaging exocytosis and endocytosis," Curr. Opin. Neurobiol. 6, 365-371 (1996).
[CrossRef] [PubMed]

Martin, E.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Maruyama, I.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Maruyamal, I.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Matsushita, Y.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Matveeva, E.

K. Aslan, I. Gryczynski, J. Malicka, E. Matveeva, J. R. Lakowicz, and C. D. Geddes, "Metal-enhanced fluorescence: an emerging tool in biotechnology," Curr. Opin. Biotechnol. 16, 55-62 (2005).
[CrossRef] [PubMed]

E. Matveeva, Z. Gryczynski, J. Malicka, I. Gryczynski, and J. R. Lakowicz, "Metal-enhanced fluorescence immunoassays using total internal reflection and silver island-coated surfaces," Anal. Biochem. 334, 303-311 (2004).
[CrossRef] [PubMed]

McDonagh, C.

O. Stranik, H. M. McEvoy, C. McDonagh, and B. D. MacCraith, "Plasmonic enhancement of fluorescence for sensor applications," Sens. Actuators B 107, 148-153 (2005).
[CrossRef]

McEvoy, H. M.

O. Stranik, H. M. McEvoy, C. McDonagh, and B. D. MacCraith, "Plasmonic enhancement of fluorescence for sensor applications," Sens. Actuators B 107, 148-153 (2005).
[CrossRef]

Nakagawa, K.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Nakagawa, M.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Natsugoe, S.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Nawroth, P. P.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Nishioka, J.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Novotny, L.

P. Anger, P. Bharadwaj, and L. Novotny, "Enhancement and quenching of single-molecule fluorescence," Phys. Rev. Lett. 96, 113002 (2006).
[CrossRef] [PubMed]

Obama, H.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Persson, B.

F. Yu, B. Persson, S. Lofas, and W. Knoll, "Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level," Anal. Chem. 76, 6765-6770 (2004).
[CrossRef] [PubMed]

Qiu, F.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Raschke, G.

G. Raschke, S. Kowarik, T. Franzl, C. So1nnichsen, T. A. Klar, J. Feldmann, A. Nichtl, and K. Ku1rzinger, "Biomolecular recognition based on single gold nanoparticle light scattering," Nano Lett. 3, 935-938 (2003).
[CrossRef]

Reichert, W. M.

G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, "Total internal reflection fluorescence microscopy (TIRFM) II. Topographical mapping of relative cell/substratum separation distances," J. Cell Sci. 103, 491-499 (1992).
[PubMed]

Rosenberg, R. D.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

Ruckstuhl, T.

Sagara, M.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Sailer, R.

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Schneckenburger, H.

H. Schneckenburger, "Total internal reflection fluorescence microscopy: technical innovations and novel applications," Curr. Opin. Biotechnol. 16, 13-18 (2005).
[CrossRef] [PubMed]

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Shi, C. S.

H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, "Thrombomodulin-mediated cell adhesion," J. Biol. Chem. 278, 46750-46759 (2003).
[CrossRef] [PubMed]

Shi, G. Y.

H. C. Huang, G. Y. Shi, S. J. Jiang, C. S. Shi, C. M. Wu, H. Y. Yang, and H. L. Wu, "Thrombomodulin-mediated cell adhesion," J. Biol. Chem. 278, 46750-46759 (2003).
[CrossRef] [PubMed]

Shimizu, T.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Shirao, K.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Smith, C. B.

W. J. Betz, F. Mao, and C. B. Smith, "Imaging exocytosis and endocytosis," Curr. Opin. Neurobiol. 6, 365-371 (1996).
[CrossRef] [PubMed]

So, C.

G. Raschke, S. Kowarik, T. Franzl, C. So1nnichsen, T. A. Klar, J. Feldmann, A. Nichtl, and K. Ku1rzinger, "Biomolecular recognition based on single gold nanoparticle light scattering," Nano Lett. 3, 935-938 (2003).
[CrossRef]

Stefani, F. D.

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

Steiner, R.

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Stern, D. M.

Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
[CrossRef] [PubMed]

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J. A. Steyer and W. Almers, "A real-time view of within life within 100 nm of the plasma membrane," Nat. Rev. Mol. Cell Biol. 2, 268-276 (2001).
[CrossRef] [PubMed]

Stock, K.

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Stoyanova, N.

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

Stranik, O.

O. Stranik, H. M. McEvoy, C. McDonagh, and B. D. MacCraith, "Plasmonic enhancement of fluorescence for sensor applications," Sens. Actuators B 107, 148-153 (2005).
[CrossRef]

Strauss, W. S.

R. Sailer, W. S. Strauss, H. Emmert, K. Stock, R. Steiner, and H. Schneckenburger, "Plasma membrane associated location of sulfonated meso-tetraphenylporphyrins of different hydrophilicity probed by total internal reflection fluorescence spectroscopy," Photochem. Photobiol. 71, 460-465 (2000).
[CrossRef] [PubMed]

Sund, S. E.

S. E. Sund and D. Axelrod, "Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleaching," Biophys. J. 79, 1655-1669 (2000).
[CrossRef] [PubMed]

Suzuki, K.

K. Suzuki, H. Kusumoto, Y. Deyashiki, J. Nishioka, I. Maruyamal, M. Zushi, S. Kawahara, G. Honda, S. Yamamoto, and S. Horiguchi, "Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation," EMBO J. 6, 1891 -1897 (1987).
[PubMed]

Tawa, K.

K. Tawa and W. Knoll, "Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy," Nucleic Acids Res. 32, 2372-2377 (2004).
[CrossRef] [PubMed]

Tezuka, Y.

Y. Tezuka, S. Yonezawa, I. Maruyama, Y. Matsushita, T. Shimizu, H. Obama, M. Sagara, K. Shirao, C. Kusano, and S. Natsugoe, "Expression of thrombomodulin in esophageal squamous cell carcinoma and its relationship to lymph node metastasis," Cancer Res. 55, 196-200 (1995).

Toomre, D.

D. Toomre and D. J. Manstein, "Lighting up the cell surface with evanescent wave microscopy," Trends Cell Biol. 11, 298-303 (2001).
[CrossRef] [PubMed]

Truskey, G. A.

G. A. Truskey, J. S. Burmeister, E. Grapa, and W. M. Reichert, "Total internal reflection fluorescence microscopy (TIRFM) II. Topographical mapping of relative cell/substratum separation distances," J. Cell Sci. 103, 491-499 (1992).
[PubMed]

Vasilev, K.

F. D. Stefani, K. Vasilev, N. Bocchio, N. Stoyanova, and M. Kreiter, "Surface-plasmon-mediated single-molecule fluorescence through a thin metallic film," Phys. Rev. Lett. 94, 023005 (2005).
[CrossRef] [PubMed]

Weber, W. H.

Wilhelm, O.

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Yu, F.

S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
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F. Yu, B. Persson, S. Lofas, and W. Knoll, "Surface plasmon fluorescence immunoassay of free prostate-specific antigen in human plasma at the femtomolar level," Anal. Chem. 76, 6765-6770 (2004).
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Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
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Colloids Surf. A

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Curr. Opin. Biotechnol.

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Y. Zhang, H. W. Guettler, J. Chen, O. Wilhelm, Y. Deng, F. Qiu, K. Nakagawa, M. Klevesath, S. Wilhelm, H. Böhrer, M. Nakagawa, H. Graeff, E. Martin, D. M. Stern, R. D. Rosenberg, R. Ziegler, and P. P. Nawroth, "Thrombomodulin modulates growth of tumor cells independent of its anticoagulant activity," J. Clin. Invest. 101, 1301-1309 (1998).
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S. Ekgasit, F. Yu, and W. Knoll, "Displacement of molecules near a metal surface as seen by an SPR-SPFS biosensor," Langmuir 21, 4077-4082 (2005).
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K. Tawa and W. Knoll, "Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy," Nucleic Acids Res. 32, 2372-2377 (2004).
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Figures (4)

Fig. 1.
Fig. 1.

(a) Schematic illustration of experimental configuration employed for live cell imaging using conventional and surface plasmon-enhanced TIRFM techniques. (b) Conventional TIR chip: cell is cultured on collagen-coated slide modified with silane. (c) Enhanced SPR chip, cell is cultured on collagen-coated silver thin film modified with thiol.

Fig. 2.
Fig. 2.

(a) Solid line: reflectivity spectrum of TIR chip as function of incident angle; Dashed line: enhancement factor of electric field intensity at interface between collagen and buffer. (b) Enhancement factor distribution is perpendicular at sensing interface at incident angle of 57.46°.

Fig. 3.
Fig. 3.

(a) Solid line: reflectivity spectrum of SPR chip as function of incident angle; Dashed line: enhancement factor of electric field intensity at interface between collagen and buffer. (b) Enhancement factor distribution is perpendicular at sensing interface at incident angle of 57.46°.

Fig. 4.
Fig. 4.

TIRFM live cell membrane images exposed at (a) 0.5 seconds and (b) 0.04 seconds; Surface plasmon-enhanced TIRFM images exposed at (c) 0.5 seconds and (d) 0.04 seconds; (e) Distribution of fluorescence intensity along x-axis of central crosscut lines of (a) conventional TIRFM live cell membrane image (dashed line) and (c) Surface plasmon-enhanced TIRFM image (solid line).

Equations (2)

Equations on this page are rendered with MathJax. Learn more.

k x = ω c ε 0 sin θ = k sp 0 = ω c ε 2 ε 1 ε 2 + ε 1 ,
R 012 = r 01 + r 12 exp ( 2 j k z 1 d 1 ) 1 + r 01 r 12 exp ( 2 j k z 1 d 1 ) 2 with r ij = ( k zi ε i k zj ε j ) ( k zi ε i + k zj ε j ) for i , j = 0 , 1 , 2 ,

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