Abstract

We describe a novel confocal microscope that uses separate low-numerical-aperture objectives with the illumination and collection axes crossed at angle θ from the midline. This architecture collects images in scattering media with high transverse and axial resolution, long working distance, large field of view, and reduced noise from scattered light. We measured transverse and axial (FWHM) resolution of 1.3 and 2.1 µm, respectively, in free space, and confirm subcellular resolution in excised esophageal mucosa. The optics may be scaled to millimeter dimensions and fiber coupled for collection of high-resolution images in vivo.

© 2003 Optical Society of America

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References

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  1. J. Pawley, ed., Handbook of Biological Confocal Microscopy, 3rd ed. (Plenum, New York, 1996).
  2. T. R. Corle and G. S. Kino, Confocal Scanning Optical Microscopy and Related Imaging Systems (Academic, Boston, Mass., 1996).
  3. M. Rajadhyaksha, R. R. Anderson, and R. H. Webb, Appl. Opt. 38, 2105 (1999).
    [CrossRef]
  4. D. L. Dickensheets and G. S. Kino, Opt. Lett. 21, 764 (1996).
    [CrossRef] [PubMed]
  5. R. H. Webb and F. Rogomentich, Appl. Opt. 38, 4870 (1999).
    [CrossRef]
  6. E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
    [CrossRef]
  7. A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
    [CrossRef] [PubMed]

2002

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

1999

1996

1994

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

Anderson, R. R.

Collier, T.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Corle, T. R.

T. R. Corle and G. S. Kino, Confocal Scanning Optical Microscopy and Related Imaging Systems (Academic, Boston, Mass., 1996).

Dickensheets, D. L.

Drezek, R.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Follen, M.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Kino, G. S.

D. L. Dickensheets and G. S. Kino, Opt. Lett. 21, 764 (1996).
[CrossRef] [PubMed]

T. R. Corle and G. S. Kino, Confocal Scanning Optical Microscopy and Related Imaging Systems (Academic, Boston, Mass., 1996).

Lindek, S.

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

Lotan, R.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Rajadhyaksha, M.

Richards-Kortum, R.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Rogomentich, F.

Stelzer, E. H. K.

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

Webb, R. H.

Zuluaga, A. F.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Appl. Opt.

J. Biomed. Opt.

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, J. Biomed. Opt. 7, 398 (2002).
[CrossRef] [PubMed]

Opt. Commun.

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

Opt. Lett.

Other

J. Pawley, ed., Handbook of Biological Confocal Microscopy, 3rd ed. (Plenum, New York, 1996).

T. R. Corle and G. S. Kino, Confocal Scanning Optical Microscopy and Related Imaging Systems (Academic, Boston, Mass., 1996).

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Figures (4)

Fig. 1
Fig. 1

Schematic diagram of the dual-axes design: D, lens diameter; other notation defined in text.

Fig. 2
Fig. 2

Schematic of the optical setup: D, detector; ADC, analog-to-digital converter; PC, personal computer; other abbreviations defined in text.

Fig. 3
Fig. 3

(a) Transverse response to chrome edge; measured 20–80% width, 1.06 µm. (b) Axial response to chrome; measured FWHM spot length, 2.1 µm.

Fig. 4
Fig. 4

This reflectance image of normal esophageal mucosa collected ex vivo reveals cell nuclei and membrane; scale bar, 20 µm.

Equations (5)

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

Δrs=0.37λn sin α0.37λnα,Δzs=0.89λn1-cos α1.78λnα2,
xs=xd cos θ±zd sin θ,    ys=yd.
Δxd=0.37λnα cos θ,    Δyd=0.37λnα,Δzd=0.37λnαsin θ.
WD=D cos θ2α.
dxEdge=0.22λnα cos θ,

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