Abstract

We present a miniature, flexible two-photon microscope consisting of a fused coherent optical fiber bundle with 30,000 cores and a gradient-index lens objective. The laser focus of a standard two-photon laser-scanning microscope was scanned over the entrance surface of the fiber bundle, resulting in sequential coupling into individual cores. Fluorescent light was detected through the fiber bundle. Micrometer-sized fluorescent beads and pollen grains were readily resolved. In addition, fluorescently labeled blood vessels were imaged through the fiber bundle in rat brain in vivo.

© 2004 Optical Society of America

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    [CrossRef] [PubMed]
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2004 (2)

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

W. Göbel, A. Nimmerjahn, and F. Helmchen, Opt. Lett. 29, 1285 (2004).
[CrossRef]

2003 (4)

2002 (3)

F. Helmchen, Exp. Physiol. 87, 6 (2002).
[CrossRef]

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

F. Helmchen, D. W. Tank, and W. Denk, Appl. Opt. 41, 2930 (2002).
[CrossRef] [PubMed]

2001 (3)

S. W. Clark, F. Ö. Ilday, and F. W. Wise, Opt. Lett. 26, 1320 (2001).
[CrossRef]

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

1999 (2)

Y. S. Sabharwal, A. R. Rouse, L. Donaldson, M. F. Hopkins, and A. F. Gmitro, Appl. Opt. 38, 7133 (1999).
[CrossRef]

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

1997 (1)

W. Denk and K. Svoboda, Neuron 18, 351 (1997).
[CrossRef] [PubMed]

Aksay, E.

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

Bird, D.

Bornhop, D. J.

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

Buess, G.

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Clark, S. W.

Denk, W.

Dickens, M. M.

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

Dombeck, D. A.

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Donaldson, L.

Dubaj, V.

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

Fee, M. S.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Gmitro, A. F.

Göbel, W.

Gu, M.

Harris, M.

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

Hasan, M. T.

Helmchen, F.

Hopkins, M. F.

Houlne, M. P.

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

Ilday, F. Ö.

Jung, J. C.

J. C. Jung and M. J. Schnitzer, Opt. Lett. 28, 902 (2003).
[CrossRef] [PubMed]

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

Kasischke, K. A.

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Knittel, J.

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Levene, M. J.

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Mazzolini, A.

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

Mehta, A. D.

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

Messerschmidt, B.

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Mitra, S.

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

Molloy, R. P.

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Nimmerjahn, A.

Omenetto, F. G.

Possner, T.

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Psaltis, D.

Rouse, A. R.

Sabharwal, Y. S.

Schnieder, L.

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Schnitzer, M. J.

J. C. Jung and M. J. Schnitzer, Opt. Lett. 28, 902 (2003).
[CrossRef] [PubMed]

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

Stepnoski, R.

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

Svoboda, K.

W. Denk and K. Svoboda, Neuron 18, 351 (1997).
[CrossRef] [PubMed]

Tank, D. W.

F. Helmchen, D. W. Tank, and W. Denk, Appl. Opt. 41, 2930 (2002).
[CrossRef] [PubMed]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Theer, P.

Tsang, M.

Webb, W. W.

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Wise, F. W.

Wood, A.

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

Appl. Opt. (2)

Exp. Physiol. (1)

F. Helmchen, Exp. Physiol. 87, 6 (2002).
[CrossRef]

J. Microsc. (1)

V. Dubaj, A. Mazzolini, A. Wood, and M. Harris, J. Microsc. 207, 108 (2002).
[CrossRef] [PubMed]

J. Neurophysiol. (1)

M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, and W. W. Webb, J. Neurophysiol. 91, 1908 (2004).
[CrossRef]

Neuron (2)

W. Denk and K. Svoboda, Neuron 18, 351 (1997).
[CrossRef] [PubMed]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, Neuron 31, 903 (2001).
[CrossRef] [PubMed]

Opt. Commun. (1)

J. Knittel, L. Schnieder, G. Buess, B. Messerschmidt, and T. Possner, Opt. Commun. 188, 267 (2001).
[CrossRef]

Opt. Eng. (1)

M. M. Dickens, M. P. Houlne, S. Mitra, and D. J. Bornhop, Opt. Eng. 38, 1836 (1999).
[CrossRef]

Opt. Lett. (6)

Other (1)

J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer, J. Neurophysiol. (to be published).

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Figures (4)

Fig. 1
Fig. 1

(a) GRIN lens objective connected to the fiber-bundle end face. Scale is in millimeters. The divergent beams emerging from the fiber cores are collimated by the imaging GRIN lens (IL) and refocused by the objective GRIN lens (OL). The optical path is exemplified for two different cores. (b) Scheme of the optical setup. Femtosecond laser pulses double pass a pair of diffraction gratings before they are coupled into the 1.4-m-long fiber bundle through a standard two-photon laser-scanning microscope. Fluorescent light is detected through the fiber bundle and the incoupling objective by a photomultiplier tube.

Fig. 2
Fig. 2

Fiber-bundle microscope images of single fluorescent beads of various sizes (top row, 6µm diameter; middle row, 3µm diameter; bottom row, 1µm diameter). Left column shows raw images, in which individual fiber-bundle cores are clearly visible. Images in the middle column were blurred with a Gaussian filter. Right column shows intensity line profiles of the raw (solid traces) and the smoothed (dashed traces) images.

Fig. 3
Fig. 3

Examples of two-photon fiber-bundle microscope images of two types of pollen grain. (a) Images taken with a standard two-photon microscope. (b) Raw images of the same types of pollen grain taken with a fiber-bundle microscope (averages of 10). (c) Smoothed visualization of the images after application of a Gaussian blur filter.

Fig. 4
Fig. 4

Fiber-bundle microscope images of Fluorescein-labeled brain microvasculature in intact neocortex of a living, anesthetized rat. Raw images (left) and smoothed images (right) are shown. The junction of two large blood vessels is visible in the upper row (average of 20). A small-caliber blood vessel is shown in the lower row (average of 10).

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