Abstract

We describe a novel technique, based on optical coherence tomography, for enhanced optical sectioning in confocal microscopy. Confocal imaging deep into highly scattering media is demonstrated and compared with the predictions of a single-backscatter theory.

© 1994 Optical Society of America

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References

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  1. T. Wilson, in Confocal Microscopy, T. Wilson, ed. (Academic, London, 1990), pp. 1–64.
    [CrossRef]
  2. C. J. Koester, in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1990), pp. 207–214.
    [CrossRef]
  3. R. C. Youngquist, S. Carr, D. E. N. Davies, Opt. Lett. 12, 158 (1987).
    [CrossRef] [PubMed]
  4. K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).
  5. E. A. Swanson, D. Huang, M. R. Hee, J. G. Fujimoto, C. P. Lin, C. A. Puliafito, Opt. Lett. 17, 151 (1992).
    [CrossRef] [PubMed]
  6. D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
    [CrossRef] [PubMed]
  7. M. R. Hee, J. A. Izatt, J. M. Jacobson, J. G. Fujimoto, E. A. Swanson, Opt. Lett. 18, 950 (1993).
    [CrossRef] [PubMed]
  8. E. A. Swanson, J. A. Izatt, M. R. Hee, D. Huang, C. P. Lin, J. S. Schuman, C. A. Puliafito, J. G. Fujimoto, Opt. Lett. 18, 1864 (1993).
    [CrossRef] [PubMed]
  9. S. Kimura, T. Wilson, Appl. Opt. 30, 2143 (1991).
    [CrossRef] [PubMed]
  10. L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
    [CrossRef]
  11. T. Sawatari, Appl. Opt. 12, 2768 (1973).
    [CrossRef] [PubMed]
  12. S. R. Chinn, E. A. Swanson, Electron. Lett. 29, 2025 (1993).
    [CrossRef]
  13. C. M. Sonnenschein, F. A. Horrigan, Appl. Opt. 10, 1600 (1971).
    [CrossRef] [PubMed]
  14. J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

1993 (4)

1992 (1)

1991 (3)

L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
[CrossRef]

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

S. Kimura, T. Wilson, Appl. Opt. 30, 2143 (1991).
[CrossRef] [PubMed]

1987 (2)

K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).

R. C. Youngquist, S. Carr, D. E. N. Davies, Opt. Lett. 12, 158 (1987).
[CrossRef] [PubMed]

1973 (1)

1971 (1)

Bonner, R. F.

J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

Carr, S.

Chang, W.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Chida, K.

K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).

Chinn, S. R.

S. R. Chinn, E. A. Swanson, Electron. Lett. 29, 2025 (1993).
[CrossRef]

Davies, D. E. N.

Flotte, T.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Fujimoto, J. G.

Gandjbakhche, A.

J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

Gregory, K.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Guniunas, L.

L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
[CrossRef]

Hee, M. R.

Horrigan, F. A.

Huang, D.

Izatt, J. A.

Jacobson, J. M.

Juskaitis, R.

L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
[CrossRef]

Kimura, S.

Knuttel, A

J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

Koester, C. J.

C. J. Koester, in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1990), pp. 207–214.
[CrossRef]

Lin, C. P.

Noda, J.

K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).

Puliafito, C. A.

Sawatari, T.

Schmitt, J. M.

J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

Schuman, J. S.

E. A. Swanson, J. A. Izatt, M. R. Hee, D. Huang, C. P. Lin, J. S. Schuman, C. A. Puliafito, J. G. Fujimoto, Opt. Lett. 18, 1864 (1993).
[CrossRef] [PubMed]

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Shatalin, S. V.

L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
[CrossRef]

Sonnenschein, C. M.

Stinson, W. G.

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Swanson, E. A.

Takada, K.

K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).

Wilson, T.

S. Kimura, T. Wilson, Appl. Opt. 30, 2143 (1991).
[CrossRef] [PubMed]

T. Wilson, in Confocal Microscopy, T. Wilson, ed. (Academic, London, 1990), pp. 1–64.
[CrossRef]

Yokohama, I.

K. Takada, I. Yokohama, K. Chida, J. Noda, Appl. Opt. 26, 1063 (1987).

Youngquist, R. C.

Appl. Opt. (4)

Electron. Lett. (2)

L. Guniunas, R. Juskaitis, S. V. Shatalin, Electron. Lett. 27, 724 (1991).
[CrossRef]

S. R. Chinn, E. A. Swanson, Electron. Lett. 29, 2025 (1993).
[CrossRef]

Opt. Lett. (4)

Proc. Soc. Photo-Opt. Instrum. Eng. (1)

J. M. Schmitt, A Knuttel, A. Gandjbakhche, R. F. Bonner, Proc. Soc. Photo-Opt. Instrum. Eng. 1889, 197 (1993).

Science (1)

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, J. G. Fujimoto, Science 254, 1178 (1991).
[CrossRef] [PubMed]

Other (2)

T. Wilson, in Confocal Microscopy, T. Wilson, ed. (Academic, London, 1990), pp. 1–64.
[CrossRef]

C. J. Koester, in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Plenum, New York, 1990), pp. 207–214.
[CrossRef]

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Figures (5)

Fig. 1
Fig. 1

Schematic of the optical coherence microscope and the model geometry: PZT, piezoelectric transducer; A/D, analog-to-digital converter.

Fig. 2
Fig. 2

Experimental axial point-spread function with a 20× microscope objective (N.A. = 0.4) obtained by scanning a mirror through the focal plane with and without coherence gating.

Fig. 3
Fig. 3

Plots of heterodyne backscatter signal normalized to the measured reflection from a mirror versus depth in model scattering media (μs ~ 6 mm−1). Plots are shown for different focal-plane depths of a 20× microscope objective. Dashed curves, predictions of the single-backscatter model.

Fig. 4
Fig. 4

Two-dimensional images of absorbing bars embedded 700 μm deep in a scattering medium with scattering coefficient μs ~ 6 mm−1. Top left, confocal image without coherence gating (electronic noise floor −64.4 dB; image mean −50.8 dB). Bottom left, OCM (shot-noise-limited noise floor −100.8 dB). Right, plots of backscatter versus depth at positions both between (A) and over (B) the absorbing bars.

Fig. 5
Fig. 5

Parametric plot of theoretical limits to OCM, assuming D = 3.7 mm, n0 = 1.33, Lc, = 20 μm, λ = 800 nm, μb = 0.3191 μs, and E = 130 mJ.

Equations (6)

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i s η e h ν P 0 2 R s ( z ) ,
R s ( z ) = μ b π D 2 L c exp ( - 2 μ t z ) 4 ( n 0 z ) 2 [ 1 + ( π D 2 4 λ n 0 z ) 2 ( 1 - n 0 z n 0 2 f ) 2 ] ,
n 0 z a + n 0 z ,
f a + n 0 ( f - a ) [ n 0 2 ( D 2 4 f 2 + 1 ) - D 2 4 f 2 ] 1 / 2 .
μ t z 1 1 2 ln [ π 2 D 2 4 n 0 2 λ 2 ( N . A . ) 2 ] = 1 2 ln ( π 2 4 n 0 2 M 2 ) ,
μ t z 2 1 2 ln [ E 2 h ν π μ b L c n 0 2 ( N . A . ) 2 ] ,

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