Abstract

We have developed a new coherent anti-Stokes Raman scattering (CARS) microscopy system with a collinear configuration for use in the fingerprint region. The system consists of a picosecond laser system and a transmission-type laser scanning microscope without a pinhole in front of the detector. The observable Raman-shift region is 9001750 cm-1, the spectral resolution is 30 cm-1, and the spatial resolution is smaller than 1 µm in the lateral direction and 3.2 µm in the depth direction, with objectives with a numerical aperture of 0.65. CARS spectra and images of polystyrene beads are demonstrated, and CARS imaging of a viable yeast cell is attempted.

© 2000 Optical Society of America

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  1. W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).
  2. M. D. Duncan, Opt. Commun. 50, 307 (1984).
    [CrossRef]
  3. M. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Lett. 7, 350 (1982).
    [CrossRef] [PubMed]
  4. A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
    [CrossRef]
  5. G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
    [CrossRef] [PubMed]
  6. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
    [CrossRef] [PubMed]
  7. M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
    [CrossRef]
  8. M. D. Levenson and S. Kano, Introduction to Nonlinear Laser Spectroscopy (Academic, Orlando, Fla., 1988).
  9. Y. R. Shen, Principles of Nonlinear Optics (Wiley, New York, 1984).

2000 (1)

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

1999 (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

1990 (2)

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

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

1985 (1)

W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).

1984 (1)

M. D. Duncan, Opt. Commun. 50, 307 (1984).
[CrossRef]

1982 (1)

Arndt-Jovin, D. J.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Brakenhoff, G. J.

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

de Lange, C. A.

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

de Mul, F. F. M.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Denk, W.

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

Duncan, M. D.

Duncan, W. D.

W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).

Greve, J.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Holtom, G. R.

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Jovin, T. M.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Kano, S.

M. D. Levenson and S. Kano, Introduction to Nonlinear Laser Spectroscopy (Academic, Orlando, Fla., 1988).

Levenson, M. D.

M. D. Levenson and S. Kano, Introduction to Nonlinear Laser Spectroscopy (Academic, Orlando, Fla., 1988).

Manuccia, T. J.

W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).

M. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Lett. 7, 350 (1982).
[CrossRef] [PubMed]

Muller, M.

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

Otto, C.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Puppels, G. J.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Reintjes, J.

W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).

M. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Lett. 7, 350 (1982).
[CrossRef] [PubMed]

Robert-Nicoud, M.

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Shen, Y. R.

Y. R. Shen, Principles of Nonlinear Optics (Wiley, New York, 1984).

Squier, J.

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

Strickler, J. H.

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

Webb, W. W.

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

Xie, X. S.

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Zumbusch, A.

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

J. Microsc. (Oxford) (1)

M. Muller, J. Squier, C. A. de Lange, and G. J. Brakenhoff, J. Microsc. (Oxford) 197, 150 (2000).
[CrossRef]

Nature (1)

G. J. Puppels, F. F. M. de Mul, C. Otto, J. Greve, M. Robert-Nicoud, D. J. Arndt-Jovin, and T. M. Jovin, Nature 347, 301 (1990).
[CrossRef] [PubMed]

Opt. Commun. (1)

M. D. Duncan, Opt. Commun. 50, 307 (1984).
[CrossRef]

Opt. Eng. (1)

W. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Eng. 24, 352 (1985).

Opt. Lett. (1)

Phys. Rev. Lett. (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Science (1)

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

Other (2)

M. D. Levenson and S. Kano, Introduction to Nonlinear Laser Spectroscopy (Academic, Orlando, Fla., 1988).

Y. R. Shen, Principles of Nonlinear Optics (Wiley, New York, 1984).

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Figures (5)

Fig. 1
Fig. 1

Schematic of the CARS microscopy system: PZT, piezoelectric transducer. See text for other definitions.

Fig. 2
Fig. 2

CARS spectra of polystyrene beads, obtained by (squares) CARS microscope and (solid curve) conventional Raman microscope.

Fig. 3
Fig. 3

(a) Relationship between the intensity of the observed signal and (circles) ω1 beam intensity. (b) Relationship between the intensity of the observed signal and (triangles) ω2 beam intensity. The solid lines are the results of curve fitting on the log–log plots. In (b), the dashed line has a slope of 1.

Fig. 4
Fig. 4

Estimation of the z-axis resolution of the CARS microscopy system. Open circles, observed data; solid curve result of curve fitting; dashed curve, differential of the solid curve.

Fig. 5
Fig. 5

CARS images in the fingerprint region: (a) polystyrene beads (diameter, 1.0 µm) in water at a 1030cm-1 Raman shift, (b) polystyrene beads at 980 cm-1, (c) yeast cell in water at 1215 cm-1.

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