Abstract

A high-resolution fluorescence microscopy technique has been developed that achieves a lateral resolution of better than one sixth of the emission wavelength (FWHM). By use of a total-internal-reflection geometry, standing evanescent waves are generated that spatially modulate the excitation of the sample. An enhanced two-dimensional image is formed from a weighted sum of images taken at different phases and directions of the standing wave. The performance of such a system is examined through theoretical calculations of both the point-spread function and the optical transfer function.

© 2000 Optical Society of America

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  1. M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
    [CrossRef]
  2. T. Wilson and C. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic, New York, 1984).
  3. E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
    [CrossRef] [PubMed]
  4. G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
    [CrossRef] [PubMed]
  5. B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
    [CrossRef] [PubMed]
  6. V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
    [CrossRef]
  7. D. Axelrod, Annu. Rev. Biophys. Bioeng. 13, 247 (1984).
    [CrossRef]
  8. J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
    [CrossRef] [PubMed]
  9. P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
    [CrossRef]
  10. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  11. S. W. Hell and J. Wichmann, Opt. Lett. 19, 780 (1994).
    [CrossRef] [PubMed]
  12. C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
    [CrossRef] [PubMed]
  13. T. A. Klar and S. W. Hell, Opt. Lett. 24, 954 (1999).
    [CrossRef]
  14. J. W. Goodman, Introduction to Fourier Optics (McGraw-Hill, New York, 1996), p. 143.

1999 (1)

1998 (1)

M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
[CrossRef]

1996 (1)

V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
[CrossRef]

1995 (2)

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

1994 (1)

1993 (1)

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

1992 (1)

J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
[CrossRef] [PubMed]

1991 (1)

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

1990 (1)

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

1986 (1)

G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
[CrossRef] [PubMed]

1984 (1)

D. Axelrod, Annu. Rev. Biophys. Bioeng. 13, 247 (1984).
[CrossRef]

Abney, J. R.

J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
[CrossRef] [PubMed]

Axelrod, D.

D. Axelrod, Annu. Rev. Biophys. Bioeng. 13, 247 (1984).
[CrossRef]

Bailey, B.

V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
[CrossRef]

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

Betzig, E.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

Binnig, G.

G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
[CrossRef] [PubMed]

Denk, W.

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

Dong, C. Y.

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

Farkas, D. L.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

French, T.

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

Gerber, Ch.

G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
[CrossRef] [PubMed]

Goodman, J. W.

J. W. Goodman, Introduction to Fourier Optics (McGraw-Hill, New York, 1996), p. 143.

Gratton, E.

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

Hanninen, P. E.

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

Harris, T. D.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

Hell, S. W.

T. A. Klar and S. W. Hell, Opt. Lett. 24, 954 (1999).
[CrossRef]

M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
[CrossRef]

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

S. W. Hell and J. Wichmann, Opt. Lett. 19, 780 (1994).
[CrossRef] [PubMed]

Klar, T. A.

Kostelak, R. L.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

Krishnamurthi, V.

V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
[CrossRef]

Lanni, F.

V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
[CrossRef]

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

Quate, C. F.

G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
[CrossRef] [PubMed]

Salo, J.

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

Scalettar, B. A.

J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
[CrossRef] [PubMed]

Schrader, M.

M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
[CrossRef]

Sheppard, C.

T. Wilson and C. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic, New York, 1984).

So, P. T. C.

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

Soini, E.

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

Strickler, J. H.

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

Taylor, D. L.

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

Thompson, N. L.

J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
[CrossRef] [PubMed]

Trautman, J. K.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

van der Voort, H. T. M.

M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
[CrossRef]

Webb, W. W.

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

Weiner, J. S.

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

Wichmann, J.

Wilson, T.

T. Wilson and C. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic, New York, 1984).

Annu. Rev. Biophys. Bioeng. (1)

D. Axelrod, Annu. Rev. Biophys. Bioeng. 13, 247 (1984).
[CrossRef]

Appl. Phys. Lett. (1)

P. E. Hanninen, S. W. Hell, J. Salo, and E. Soini, Appl. Phys. Lett. 66, 1698 (1995).
[CrossRef]

Biophys. J. (2)

C. Y. Dong, P. T. C. So, T. French, and E. Gratton, Biophys. J. 69, 2234 (1995).
[CrossRef] [PubMed]

J. R. Abney, B. A. Scalettar, and N. L. Thompson, Biophys. J. 61, 542 (1992).
[CrossRef] [PubMed]

J. Appl. Phys. (1)

M. Schrader, S. W. Hell, and H. T. M. van der Voort, J. Appl. Phys. 84, 4033 (1998).
[CrossRef]

Nature (1)

B. Bailey, D. L. Farkas, D. L. Taylor, and F. Lanni, Nature 366, 44 (1993).
[CrossRef] [PubMed]

Opt. Lett. (2)

Phys. Rev. Lett. (1)

G. Binnig, C. F. Quate, and Ch. Gerber, Phys. Rev. Lett. 56, 930 (1986).
[CrossRef] [PubMed]

Proc. SPIE (1)

V. Krishnamurthi, B. Bailey, and F. Lanni, Proc. SPIE 2655, 18 (1996).
[CrossRef]

Science (2)

E. Betzig, J. K. Trautman, T. D. Harris, J. S. Weiner, and R. L. Kostelak, Science 251, 1468 (1991).
[CrossRef] [PubMed]

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

Other (2)

J. W. Goodman, Introduction to Fourier Optics (McGraw-Hill, New York, 1996), p. 143.

T. Wilson and C. Sheppard, Theory and Practice of Scanning Optical Microscopy (Academic, New York, 1984).

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Figures (2)

Fig. 1
Fig. 1

For the representative parameters of N=4, θi=75°, NA=1.3, λ=532 nm, and λe=560 nm, plots are generated that show (a) the PSF’s of conventional, confocal, and SWTIRF microscopies, (b) the PSF of a one-dimensional (1D) SWTIRF (quartz, n=1.46) scan [Eq. (10)], (c) the symmetric PSF of two-dimensional (2D) SWTIRF (quartz) generated by Eq. (14).

Fig. 2
Fig. 2

(a) and (b), Frequency space counterparts (OTF’s) of Figs. 1(b) and 1(c), respectively.

Equations (15)

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

Ispr=Or1+cosKx+ϕ,
K=4πn sinθi/λ,
I0r=Or1+cosKx+ϕPr,
Ijr=OrPr+1/2expiKx+ϕ+jπ/2OrPr+1/2exp-iKx+ϕ+jπ/2OrPr,   j=0,1,2,
OrPr=1/2I0r+I2r,
1/2expiKx+ϕOrPr=1/41-iI0r+2iI1r-1+iI2r,
1/2exp-iKx+ϕOrPr=1/41+iI0r-2iI1r+-1+iI2r.
exp±iKx+ϕOrPr=exp±iKx+ϕOrexpiKxPr,
Ir=OrPr1+cosKx-ϕ+α=I0r1/21+2cosKx+α+π/4+I1rsinKx+α+I2r1/2×1-2cosKx+α-π/4.
PSWxr=1+cosKxPr.
P~SWxk=2+P~-Kex+P~Kex-1×2P~k+P~k-Kex+P~k+Kex,
PSWξr,ξ=1+cosKr cosθ-ξPr,
PSWr,N=11-β1Nm=0N-11+cosKr cosθ-mπ/N-βPr=11-βSr,NPr,
PSWr,N=1.431Nm=0N-11+cosKr cosθ-mπ/N-0.3Pr;
P˜SWk,N=γ1Nm=02N-1P˜k+Kcosmπ/Nex+sinmπ/Ney+1.7P˜k,

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