Abstract

We present a demonstration of near-field scanning optical microscopy of single molecules based on ultrafast two-photon-induced fluorescence. Measurements were performed by use of 100-fs pulses at 800  nm from a Ti:sapphire laser to excite the two-photon transition in Rhodamine B molecules. Although near-field probes are normally metal coated to achieve superresolution, we used uncoated tips to achieve sufficiently high optical powers to generate acceptable fluorescence emission rates. Images of single molecules demonstrate a resolution of 175 nm <λ/4 on a topographically smooth surface, which surpasses the apparent λ/2 resolution limit for uncoated tips operating in the linear response regime.

© 1998 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
    [CrossRef]
  2. A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
    [CrossRef]
  3. M. A. Paesler and P. J. Moyer, Near-Field Optics (Wiley, New York, 1996), p. 288.
  4. R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
    [CrossRef] [PubMed]
  5. E. Betzig and R. J. Chichester, Science 262, 1422 (1993); X. S. Xie and R. C. Dunn, Science 265, 361 (1994); W. P. Ambrose, P. M. Goodwin, J. C. Martin, and R. A. Keller, Science 265, 364 (1994).
    [CrossRef] [PubMed]
  6. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  7. B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
    [CrossRef]
  8. V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
    [CrossRef]
  9. C. Xu and W. W. Web, J. Opt. Soc. Am. B 13, 481 (1996).
    [CrossRef]
  10. E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
    [CrossRef]
  11. R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
    [CrossRef]
  12. E. B. Treacy, IEEE J. Quantum Electron. QE-5, 454 (1969).
    [CrossRef]
  13. E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
    [CrossRef]
  14. L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
    [CrossRef]
  15. E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
    [CrossRef]
  16. L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
    [CrossRef]

1997 (4)

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

1996 (1)

1995 (2)

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
[CrossRef]

1993 (2)

E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
[CrossRef]

E. Betzig and R. J. Chichester, Science 262, 1422 (1993); X. S. Xie and R. C. Dunn, Science 265, 361 (1994); W. P. Ambrose, P. M. Goodwin, J. C. Martin, and R. A. Keller, Science 265, 364 (1994).
[CrossRef] [PubMed]

1992 (2)

E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
[CrossRef]

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

1984 (2)

D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
[CrossRef]

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

1969 (1)

E. B. Treacy, IEEE J. Quantum Electron. QE-5, 454 (1969).
[CrossRef]

Allen, E. V.

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Anderson, G. A.

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Betzig, E.

E. Betzig and R. J. Chichester, Science 262, 1422 (1993); X. S. Xie and R. C. Dunn, Science 265, 361 (1994); W. P. Ambrose, P. M. Goodwin, J. C. Martin, and R. A. Keller, Science 265, 364 (1994).
[CrossRef] [PubMed]

E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
[CrossRef]

Bielefeldt, H.

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

Brand, L.

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Buckland, E. L.

E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
[CrossRef]

Chen, Y.

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

Chichester, R. J.

E. Betzig and R. J. Chichester, Science 262, 1422 (1993); X. S. Xie and R. C. Dunn, Science 265, 361 (1994); W. P. Ambrose, P. M. Goodwin, J. C. Martin, and R. A. Keller, Science 265, 364 (1994).
[CrossRef] [PubMed]

Denk, W.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
[CrossRef]

Drexhage, K. H.

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Dunn, R. C.

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Eggeling, C.

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Finn, P. L.

E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
[CrossRef]

Harootunian, A.

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

Hecht, B.

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
[CrossRef]

Holtom, G. R.

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

Inouye, Y.

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

Isaacson, M.

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

Joyce, S. A.

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Krausch, G.

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

Lanz, M.

D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
[CrossRef]

Lewis, A.

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

Mlynek, J.

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

Moyer, P. J.

E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
[CrossRef]

M. A. Paesler and P. J. Moyer, Near-Field Optics (Wiley, New York, 1996), p. 288.

Muray, A.

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

Novotny, L.

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
[CrossRef]

Paesler, M. A.

E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
[CrossRef]

M. A. Paesler and P. J. Moyer, Near-Field Optics (Wiley, New York, 1996), p. 288.

Pohl, D. W.

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
[CrossRef]

D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
[CrossRef]

Sanchez, E. J.

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

Sandoghdar, V.

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

Seidel, C. A. M.

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Strickler, J. H.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

Toledo-Crow, R.

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

Treacy, E. B.

E. B. Treacy, IEEE J. Quantum Electron. QE-5, 454 (1969).
[CrossRef]

Vaez-Iravani, M.

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

Web, W. W.

Webb, W. W.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

Wegscheider, S.

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

Weiner, J. S.

E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
[CrossRef]

Xie, X. S.

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Xu, C.

Yang, P. C.

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

Zander, C.

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Appl. Phys. Lett. (3)

D. W. Pohl, W. Denk, and M. Lanz, Appl. Phys. Lett. 44, 651 (1984).
[CrossRef]

E. Betzig, P. L. Finn, and J. S. Weiner, Appl. Phys. Lett. 60, 2484 (1992).
[CrossRef]

R. Toledo-Crow, P. C. Yang, Y. Chen, and M. Vaez-Iravani, Appl. Phys. Lett. 60, 2957 (1992).
[CrossRef]

IEEE J. Quantum Electron. (1)

E. B. Treacy, IEEE J. Quantum Electron. QE-5, 454 (1969).
[CrossRef]

J. Appl. Phys. (3)

E. L. Buckland, P. J. Moyer, and M. A. Paesler, J. Appl. Phys. 73, 1018 (1993).
[CrossRef]

B. Hecht, H. Bielefeldt, Y. Inouye, and D. W. Pohl, J. Appl. Phys. 81, 2492 (1997).
[CrossRef]

V. Sandoghdar, S. Wegscheider, G. Krausch, and J. Mlynek, J. Appl. Phys. 81, 2499 (1997).
[CrossRef]

J. Opt. Soc. Am. B (1)

J. Phys. Chem. A (2)

E. J. Sanchez, L. Novotny, G. R. Holtom, and X. S. Xie, J. Phys. Chem. A 101, 7019 (1997).
[CrossRef]

L. Brand, C. Eggeling, C. Zander, K. H. Drexhage, and C. A. M. Seidel, J. Phys. Chem. A 101, 4313 (1997).
[CrossRef]

Science (2)

E. Betzig and R. J. Chichester, Science 262, 1422 (1993); X. S. Xie and R. C. Dunn, Science 265, 361 (1994); W. P. Ambrose, P. M. Goodwin, J. C. Martin, and R. A. Keller, Science 265, 364 (1994).
[CrossRef] [PubMed]

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
[CrossRef] [PubMed]

Ultramicroscopy (3)

A. Lewis, M. Isaacson, A. Harootunian, and A. Muray, Ultramicroscopy 13, 227 (1984).
[CrossRef]

L. Novotny, D. W. Pohl, and B. Hecht, Ultramicroscopy 61, 1 (1995).
[CrossRef]

R. C. Dunn, E. V. Allen, S. A. Joyce, G. A. Anderson, and X. S. Xie, Ultramicroscopy 57, 113 (1995); J. Hwang, L. K. Tamm, C. Bohm, T. S. Ramalingam, E. Betzig, and M. Edidin, Science 270, 610 (1995); T. Enderle, T. Ha, D. F. Ogletree, D. S. Chemla, C. Magowan, and S. Weiss, Proc. Natl. Acad. Sci. U.S.A. 94, 520 (1997).
[CrossRef] [PubMed]

Other (1)

M. A. Paesler and P. J. Moyer, Near-Field Optics (Wiley, New York, 1996), p. 288.

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (3)

Fig. 1
Fig. 1

Schematic of the two-photon near-field scanning optical microscopy (TP-NSOM) system. Single dye molecules were excited by light from a mode-locked Ti:sapphire laser transmitted through an uncoated probe. An oil-immersion objective and a bandpass filter collected and spectrally isolated the resulting fluorescence, which was detected by an avalanche photodiode (APD). A four-quadrant piezo tube scanned the sample.

Fig. 2
Fig. 2

Image of two-photon-induced fluorescence from single Rhodamine B molecules. Each peak represents the fluorescence from a single molecule. The maximum signal in the image is 180 counts. The molecules were excited by light from a mode-locked 100-fs Ti:sapphire laser operating at 800  nm with a repetition rate of 80  MHz and an average transmitted power of 150 µW. The image was smoothed by use of a weighted nearest-neighbor average to remove noise fluctuations.

Fig. 3
Fig. 3

Line scan through the fluorescence image of a single molecule. The raw data are well fitted by a Gaussian function with a FWHM of 160  nm. The residuals of the fit are also displayed.

Metrics