Abstract

We report a scheme for 2D standing-wave total-internal-reflection fluorescence microscopy. Standing-wave patterns are generated by two interfering beams coupled through the objective lens. Period, angular orientation, and phase of standing waves are controlled entirely by acousto-optic deflectors. The lateral resolution improvement of 100nm is combined with an axial selectivity of <100nm by utilizing an evanescent standing-wave pattern. This technique can provide real-time imaging of subresolution structures in live biological specimens near a glass–water interface.

© 2009 Optical Society of America

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References

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    [CrossRef] [PubMed]
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2008 (3)

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

M. Beck, M. Aschwanden, and A. Stemmer, J. Microsc. 232, 99 (2008).
[CrossRef] [PubMed]

R. Fiolka, M. Beck, and A. Stemmer, Opt. Lett. 33, 1629 (2008).
[CrossRef] [PubMed]

2007 (1)

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

2006 (2)

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

E. Chung, D. Kim, and P. T. C. So, Opt. Lett. 31, 945 (2006).
[CrossRef] [PubMed]

2005 (2)

M. G. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

V. Westphal and S. W. Hell, Phys. Rev. Lett. 94, 143903 (2005).
[CrossRef] [PubMed]

2000 (2)

M. G. Gustafsson, J. Microsc. 198, 82 (2000).
[CrossRef] [PubMed]

J. T. Frohn, H. F. Knapp, and A. Stemmer, Proc. Natl. Acad. Sci. USA 97, 7232 (2000).
[CrossRef] [PubMed]

1999 (1)

R. Heintzmann and C. Cremer, Proc. SPIE 3568, 185 (1999).
[CrossRef]

1981 (1)

D. Axelrod, J. Cell Biol. 89, 141 (1981).
[CrossRef] [PubMed]

Agard, D. A.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Anvari, B.

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

Aschwanden, M.

M. Beck, M. Aschwanden, and A. Stemmer, J. Microsc. 232, 99 (2008).
[CrossRef] [PubMed]

Axelrod, D.

D. Axelrod, J. Cell Biol. 89, 141 (1981).
[CrossRef] [PubMed]

Beck, M.

Brownell, W. E.

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

Cande, W. Z.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Carlton, P. M.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Chung, E.

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

E. Chung, D. Kim, and P. T. C. So, Opt. Lett. 31, 945 (2006).
[CrossRef] [PubMed]

Cremer, C.

R. Heintzmann and C. Cremer, Proc. SPIE 3568, 185 (1999).
[CrossRef]

Cui, Y.

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

Fiolka, R.

Frohn, J. T.

J. T. Frohn, H. F. Knapp, and A. Stemmer, Proc. Natl. Acad. Sci. USA 97, 7232 (2000).
[CrossRef] [PubMed]

Gliko, O.

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

Golubovskaya, I. N.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Gustafsson, M. G.

M. G. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

M. G. Gustafsson, J. Microsc. 198, 82 (2000).
[CrossRef] [PubMed]

Gustafsson, M. G. L.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Heintzmann, R.

R. Heintzmann and C. Cremer, Proc. SPIE 3568, 185 (1999).
[CrossRef]

Hell, S. W.

V. Westphal and S. W. Hell, Phys. Rev. Lett. 94, 143903 (2005).
[CrossRef] [PubMed]

Kim, D.

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

E. Chung, D. Kim, and P. T. C. So, Opt. Lett. 31, 945 (2006).
[CrossRef] [PubMed]

Kim, Y. H.

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

Knapp, H. F.

J. T. Frohn, H. F. Knapp, and A. Stemmer, Proc. Natl. Acad. Sci. USA 97, 7232 (2000).
[CrossRef] [PubMed]

Reddy, G. D.

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

Saggau, P.

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

Sedat, J. W.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Shao, L.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

So, P. T. C.

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

E. Chung, D. Kim, and P. T. C. So, Opt. Lett. 31, 945 (2006).
[CrossRef] [PubMed]

Stemmer, A.

R. Fiolka, M. Beck, and A. Stemmer, Opt. Lett. 33, 1629 (2008).
[CrossRef] [PubMed]

M. Beck, M. Aschwanden, and A. Stemmer, J. Microsc. 232, 99 (2008).
[CrossRef] [PubMed]

J. T. Frohn, H. F. Knapp, and A. Stemmer, Proc. Natl. Acad. Sci. USA 97, 7232 (2000).
[CrossRef] [PubMed]

Stroud, R.

J. Xu and R. Stroud, Acousto-Optic Devices: Principles, Design, and Applications (Wiley, 1992).

Wang, C. J. R.

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

Westphal, V.

V. Westphal and S. W. Hell, Phys. Rev. Lett. 94, 143903 (2005).
[CrossRef] [PubMed]

Xu, J.

J. Xu and R. Stroud, Acousto-Optic Devices: Principles, Design, and Applications (Wiley, 1992).

Biophys. J. (2)

M. G. L. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, Biophys. J. 94, 4957 (2008).
[CrossRef] [PubMed]

E. Chung, D. Kim, Y. Cui, Y. H. Kim, and P. T. C. So, Biophys. J. 93, 1747 (2007).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

O. Gliko, G. D. Reddy, B. Anvari, W. E. Brownell, and P. Saggau, J. Biomed. Opt. 11, 064013 (2006).
[CrossRef]

J. Cell Biol. (1)

D. Axelrod, J. Cell Biol. 89, 141 (1981).
[CrossRef] [PubMed]

J. Microsc. (2)

M. Beck, M. Aschwanden, and A. Stemmer, J. Microsc. 232, 99 (2008).
[CrossRef] [PubMed]

M. G. Gustafsson, J. Microsc. 198, 82 (2000).
[CrossRef] [PubMed]

Opt. Lett. (2)

Phys. Rev. Lett. (1)

V. Westphal and S. W. Hell, Phys. Rev. Lett. 94, 143903 (2005).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. USA (2)

J. T. Frohn, H. F. Knapp, and A. Stemmer, Proc. Natl. Acad. Sci. USA 97, 7232 (2000).
[CrossRef] [PubMed]

M. G. Gustafsson, Proc. Natl. Acad. Sci. USA 102, 13081 (2005).
[CrossRef] [PubMed]

Proc. SPIE (1)

R. Heintzmann and C. Cremer, Proc. SPIE 3568, 185 (1999).
[CrossRef]

Other (2)

J.B.Pawley, ed., Handbook of Biological Confocal Microscopy (Springer Science, 2006).
[CrossRef]

J. Xu and R. Stroud, Acousto-Optic Devices: Principles, Design, and Applications (Wiley, 1992).

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Figures (2)

Fig. 1
Fig. 1

2D SW-TIRFM setup. Inset 1, interfering beams at the object plane; inset 2, focused beams at the backfocal plane.

Fig. 2
Fig. 2

Imaging 100 nm fluorescent beads using SW-TIRFM: (a) TIRFM, (b) deconvolved TIRFM, and (c) SW-TIRFM images; (d) intensity profiles of images (b) and (c) along the x and y directions through the central bead indicate 2D improved resolution of 100 nm and highly isotropic effective PSF.

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