Abstract

We introduce 4Pi confocal microscopy with destructive interference of converging waves. Linear lobe deconvolution as well as nonlinear restoration of 4Pi confocal raw data with their point-spread functions (PSF’s) leads to almost-identical images, irrespective of whether the 4Pi confocal PSF relies on constructive or destructive interference. Three-dimensional imaging of microtubules of a mouse fibroblast cell yielded an axial resolution near 100-nm in both cases. Moreover, restoration of 4Pi confocal images of the same object with alternate phases is introduced as a powerful test for (nonlinear) image restoration.

© 1998 Optical Society of America

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  1. S. Hell, “Double-confocal scanning microscope,” European patent0491289 (December18, 1996); S. Hell and E. H. K. Stelzer, J. Opt. Soc. Am. A 9, 2159 (1992); Opt. Commun. 93, 277 (1992); M. Schrader and S. W. Hell, J. Microsc. 183, 189 (1996); M. Schrader, M. Kozubek, S. W. Hell, and T. Wilson, Opt. Lett. 22, 436 (1997).
    [CrossRef] [PubMed]
  2. S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
    [CrossRef]
  3. P. E. Hänninen, S. W. Hell, J. Salo, E. Soini, and C. Cremer, Appl. Phys. Lett. 66, 1698 (1995).
    [CrossRef]
  4. W. H. Richardson, J. Opt. Soc. Am 62, 55 (1972); L. B. Lucy, Astron. J. 79, 745 (1974).
    [CrossRef]
  5. W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
    [CrossRef] [PubMed]
  6. M. Gu and C. J. R. Sheppard, Opt. Commun. 114, 45 (1995).
    [CrossRef]

1997 (1)

S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
[CrossRef]

1995 (3)

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

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

M. Gu and C. J. R. Sheppard, Opt. Commun. 114, 45 (1995).
[CrossRef]

1972 (1)

W. H. Richardson, J. Opt. Soc. Am 62, 55 (1972); L. B. Lucy, Astron. J. 79, 745 (1974).
[CrossRef]

Carrington, W. A.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Cremer, C.

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

Fay, F. S.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Fogarty, K. E.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Gu, M.

M. Gu and C. J. R. Sheppard, Opt. Commun. 114, 45 (1995).
[CrossRef]

Hänninen, P. E.

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

Hell, S.

S. Hell, “Double-confocal scanning microscope,” European patent0491289 (December18, 1996); S. Hell and E. H. K. Stelzer, J. Opt. Soc. Am. A 9, 2159 (1992); Opt. Commun. 93, 277 (1992); M. Schrader and S. W. Hell, J. Microsc. 183, 189 (1996); M. Schrader, M. Kozubek, S. W. Hell, and T. Wilson, Opt. Lett. 22, 436 (1997).
[CrossRef] [PubMed]

Hell, S. W.

S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
[CrossRef]

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

Isenberg, G.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Lynch, R. M.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Moore, E. D. W.

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Richardson, W. H.

W. H. Richardson, J. Opt. Soc. Am 62, 55 (1972); L. B. Lucy, Astron. J. 79, 745 (1974).
[CrossRef]

Salo, J.

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

Schrader, M.

S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
[CrossRef]

Sheppard, C. J. R.

M. Gu and C. J. R. Sheppard, Opt. Commun. 114, 45 (1995).
[CrossRef]

Soini, E.

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

van der Voort, H. T. M.

S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
[CrossRef]

Appl. Phys. Lett. (1)

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

J. Microsc. (1)

S. W. Hell, M. Schrader, and H. T. M. van der Voort, J. Microsc. 185, 1 (1997).
[CrossRef]

J. Opt. Soc. Am (1)

W. H. Richardson, J. Opt. Soc. Am 62, 55 (1972); L. B. Lucy, Astron. J. 79, 745 (1974).
[CrossRef]

Opt. Commun. (1)

M. Gu and C. J. R. Sheppard, Opt. Commun. 114, 45 (1995).
[CrossRef]

Science (1)

W. A. Carrington, R. M. Lynch, E. D. W. Moore, G. Isenberg, K. E. Fogarty, and F. S. Fay, Science 268, 1483 (1995); G. M. P. Van Kempen, L. J. Van Vliet, P. J. Verveer, and H. T. M. van der Voort, J. Microsc. 185, 354 (1997).
[CrossRef] [PubMed]

Other (1)

S. Hell, “Double-confocal scanning microscope,” European patent0491289 (December18, 1996); S. Hell and E. H. K. Stelzer, J. Opt. Soc. Am. A 9, 2159 (1992); Opt. Commun. 93, 277 (1992); M. Schrader and S. W. Hell, J. Microsc. 183, 189 (1996); M. Schrader, M. Kozubek, S. W. Hell, and T. Wilson, Opt. Lett. 22, 436 (1997).
[CrossRef] [PubMed]

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Figures (3)

Fig. 1
Fig. 1

Measured 4Pi confocal PSF’s for (a) constructive and (b) destructive interference along with (c) and (d) the corresponding inverse filters. (e), (f) Final, effective PSF’s for constructive and destructive interference after point deconvolution with inverse filters (c) and (d), respectively.

Fig. 2
Fig. 2

Point-deconvolved 4Pi confocal xz images from the same area of the microtubule network of a mouse fibroblast cell as obtained with (a) constructive and (b) destructive interference. (c) Confocal counterpart. Note the similarity between (a) and (b) as well as the increased resolution compared with that of (c).

Fig. 3
Fig. 3

xz slices obtained with 4Pi confocal microscopy combined with image restoration. Whereas the data that make up (a) were recorded and restored with the 4Pi PSF of Fig.  1(a), (b) was independently recorded and restored with the 4Pi PSF of Fig.  1(b). The similarity between (a) and (b) indicates the reliability of 4Pi confocal imaging combined with restoration and the total resolution improvement over that of (c), the standard confocal microscope image.

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