Abstract

Single-molecule localization (SML) is a powerful tool to overcome the diffraction limit in optical imaging, because the fluorescence emitted by single molecules can be observed with nanometer accuracy when the optical background and associated noise are made sufficiently small. Random adsorbed SML has been successfully demonstrated for superresolution imaging on metal surfaces. To optimize the random adsorbed SML, we developed a new illumination method based on surface plasmon resonance (SPR). The enhancement of the fluorescence signal and the reduction of background noise were achieved simultaneously. A high localization resolution of 15nm was demonstrated with this new SPR illumination system.

© 2011 Optical Society of America

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    [CrossRef] [PubMed]
  2. 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, Science 313, 1642(2006).
    [CrossRef] [PubMed]
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    [CrossRef]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [PubMed]
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    [CrossRef]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]

2011 (1)

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

2008 (1)

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

2007 (1)

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

2006 (2)

M. J. Rust, M. Bates, and X. Zhuang, Nat. Methods 3, 793 (2006).
[CrossRef] [PubMed]

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

2005 (1)

J. R. Lakowicz, Anal. Biochem. 337, 171 (2005).
[CrossRef] [PubMed]

2002 (1)

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

2001 (1)

D. Roy, Opt. Commun. 200, 119 (2001).
[CrossRef]

1990 (1)

W. H. Reichert and G. A. Truskey, J. Cell Sci. 96, 219 (1990).
[PubMed]

1968 (1)

E. Kretschmann and H. Raether, Z. Naturforsch. A 23, 2135 (1968).

Bates, M.

M. J. Rust, M. Bates, and X. Zhuang, Nat. Methods 3, 793 (2006).
[CrossRef] [PubMed]

Betzig, E.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Bock, H.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Bonifacino, J. S.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Cang, H.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Davidson, M. W.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Eggeling, C.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Egner, A.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Geisler, C.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Gladden, C.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Hell, S. W.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Hess, H. F.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Kretschmann, E.

E. Kretschmann and H. Raether, Z. Naturforsch. A 23, 2135 (1968).

Labno, A.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Lakowicz, J. R.

J. R. Lakowicz, Anal. Biochem. 337, 171 (2005).
[CrossRef] [PubMed]

Larson, D. R.

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Lindwasser, O. W.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Lippincott-Schwartz, J.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Liu, M.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Liu, Y. M.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Liu, Z. W.

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

Lu, C. G.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Olenych, S.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Patterson, G. H.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Raether, H.

E. Kretschmann and H. Raether, Z. Naturforsch. A 23, 2135 (1968).

Reichert, W. H.

W. H. Reichert and G. A. Truskey, J. Cell Sci. 96, 219 (1990).
[PubMed]

Roy, D.

D. Roy, Opt. Commun. 200, 119 (2001).
[CrossRef]

Rust, M. J.

M. J. Rust, M. Bates, and X. Zhuang, Nat. Methods 3, 793 (2006).
[CrossRef] [PubMed]

Schöle, A.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Sougrat, R.

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Sun, C.

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

Thompson, R. E.

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Truskey, G. A.

W. H. Reichert and G. A. Truskey, J. Cell Sci. 96, 219 (1990).
[PubMed]

von Middendorff, C.

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Webb, W. W.

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Wu, D. M.

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

Yin, X. B.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Zhang, X.

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

Zhuang, X.

M. J. Rust, M. Bates, and X. Zhuang, Nat. Methods 3, 793 (2006).
[CrossRef] [PubMed]

Anal. Biochem. (1)

J. R. Lakowicz, Anal. Biochem. 337, 171 (2005).
[CrossRef] [PubMed]

Appl. Phys. A (1)

C. Geisler, A. Schöle, C. von Middendorff, H. Bock, C. Eggeling, A. Egner, and S. W. Hell, Appl. Phys. A 88, 223 (2007).
[CrossRef]

Biophys. J. (1)

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

J. Cell Sci. (1)

W. H. Reichert and G. A. Truskey, J. Cell Sci. 96, 219 (1990).
[PubMed]

Nano Lett. (1)

D. M. Wu, Z. W. Liu, C. Sun, and X. Zhang, Nano Lett. 8, 1159 (2008).
[CrossRef] [PubMed]

Nat. Methods (1)

M. J. Rust, M. Bates, and X. Zhuang, Nat. Methods 3, 793 (2006).
[CrossRef] [PubMed]

Nature (1)

H. Cang, A. Labno, C. G. Lu, X. B. Yin, M. Liu, C. Gladden, Y. M. Liu, and X. Zhang, Nature 469, 385 (2011).
[CrossRef] [PubMed]

Opt. Commun. (1)

D. Roy, Opt. Commun. 200, 119 (2001).
[CrossRef]

Science (1)

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, Science 313, 1642(2006).
[CrossRef] [PubMed]

Z. Naturforsch. A (1)

E. Kretschmann and H. Raether, Z. Naturforsch. A 23, 2135 (1968).

Supplementary Material (2)

» Media 1: MOV (496 KB)     
» Media 2: MOV (157 KB)     

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Figures (5)

Fig. 1
Fig. 1

(a) Schematic diagram of SML with random adsorbed fluorescent probes, adapted from [4]. (b) Schematic diagram of the proposed SPR illumination.

Fig. 2
Fig. 2

(a) Simulated the electric field at the PMMA/water interface based on approximate calculation of multilayer transmittivity. (b) Calculated emission rate for the fluorescing molecules, which are sparsely dispersed in the water near the Au layer. (c) Profile of | E | 2 / | E i | 2 along the z direction in the four-layer SPR system with resonant angle 69.5 ° and nonresonant angle 64.5 .

Fig. 3
Fig. 3

Picture of the experiment setup for random adsorbed SML based on the SPR illumination. The schematic diagram of the configuration is shown in Fig. 1a.

Fig. 4
Fig. 4

Dynamics of the single-molecule fluorescent images captured by the CCD. (a)–(d) are captured with incident angle 64.5 ° ; (e)–(h) are captured at the resonant angle 69.5 ° . There is a fixed reference particle with similar intensity as the probe signals (indicated by circle) in the measurements (Media 1, Media 2).

Fig. 5
Fig. 5

Histogram of a single-molecule signal at (a) non resonant angle 64.5 ° and (b) resonant angle 69.5 ° , and the position distribution of subsequent measurements of a fixed reference particle at (c) nonresonant angle 64.5 ° and (d) resonant angle 69.5 ° .

Tables (1)

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Table 1 Parameters of the Configuration

Equations (2)

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F = η σ | E ( z ) | 2 ,
γ γ 0 = 1 + 3 η n 0 2 2 μ 0 k 0 3 Im ( E R ) ,

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