Abstract

A rapid line-scan confocal imager was developed for functional imaging of the retina. In this imager, an acousto-optic deflector was employed to produce mechanical vibration- and inertia-free light scanning, and a high-speed (68,000 Hz) linear CCD camera was used to achieve subcellular and submillisecond spatiotemporal resolution imaging. Two imaging modalities, i.e., frame-by-frame and line-by-line recording, were validated for the reflected light detection of intrinsic optical signals (IOSs) in visible light stimulus activated frog retinas. Experimental results indicated that fast IOSs were tightly correlated with retinal stimuli and could track visible light flicker stimulus frequency up to at least 2 Hz.

© 2010 Optical Society of America

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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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2009 (2)

2008 (5)

Y. B. Zhao and X. C. Yao, Opt. Lett. 33, 342 (2008).
[CrossRef] [PubMed]

X. C. Yao and Y. B. Zhao, Opt. Express 16, 12446 (2008).
[CrossRef] [PubMed]

K. Grieve and A. Roorda, Invest. Ophthalmol. Visual Sci. 49, 713 (2008).
[CrossRef]

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

2007 (2)

2006 (3)

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

2005 (3)

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

X. C. Yao, A. Yamauchi, B. Perry, and J. S. George, Appl. Opt. 44, 2019 (2005).
[CrossRef] [PubMed]

K. B. Im, S. Han, H. Park, D. Kim, and B. M. Kim, Opt. Express 13, 5151 (2005).
[CrossRef] [PubMed]

2002 (1)

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

1990 (1)

M. Kahlert, D. R. Pepperberg, and K. P. Hofmann, Nature 345, 537 (1990).
[CrossRef] [PubMed]

1988 (1)

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

Abramoff, M. D.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Aguirre, A. D.

Ahnelt, P.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Aloni, E.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Anger, E.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Belkin, M.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Bigelow, C. E.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Bizheva, K.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Cense, B.

Chen, Y.

Chung, C. K.

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

Curcio, C. A.

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

Drexler, W.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Duker, J. S.

Ferguson, R. D.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Foust, A. J.

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

Fujimoto, J. G.

Gao, W.

George, J. S.

Grieve, K.

K. Grieve and A. Roorda, Invest. Ophthalmol. Visual Sci. 49, 713 (2008).
[CrossRef]

Grinvald, A.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Hammer, D. X.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Han, S.

Hanazono, G.

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

Hermann, B.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Hoang, Q. V.

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

Hofmann, K. P.

M. Kahlert, D. R. Pepperberg, and K. P. Hofmann, Nature 345, 537 (1990).
[CrossRef] [PubMed]

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

Huang, S. W.

Iftimia, N. V.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Im, K. B.

Jonnal, R. S.

Kahlert, M.

M. Kahlert, D. R. Pepperberg, and K. P. Hofmann, Nature 345, 537 (1990).
[CrossRef] [PubMed]

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

Kardon, R.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Kazato, Y.

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

Kim, B. M.

Kim, D.

Krause, A.

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

Krupsky, S.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Kwon, Y. H.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Linsenmeier, R. A.

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

McCluskey, M. D.

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

Miller, D. T.

Nelson, D. A.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Park, H.

Pepperberg, D. R.

M. Kahlert, D. R. Pepperberg, and K. P. Hofmann, Nature 345, 537 (1990).
[CrossRef] [PubMed]

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

Perry, B.

Pflug, R.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Pokorny, J.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Pollack, A.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Popov, S.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Povazay, B.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Qiu, P.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Rector, D. M.

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

Reitsamer, H.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Rha, J.

Roorda, A.

K. Grieve and A. Roorda, Invest. Ophthalmol. Visual Sci. 49, 713 (2008).
[CrossRef]

Rosner, M.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Sattmann, H.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Schei, J. L.

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

Soliz, P.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Srinivasan, V. J.

Tanifuji, M.

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

Taylor, J. R.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Ts'o, D.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Tsubota, K.

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

Tsunoda, K.

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

Unterhuber, A.

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Ustun, T. E.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Vanzetta, I.

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Webb, R. H.

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Yamauchi, A.

Yao, X. C.

Zhang, Y.

Zhao, Y. B.

Zimmerman, B.

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

Appl. Opt. (1)

Invest. Ophthalmol. Visual Sci. (3)

M. D. Abramoff, Y. H. Kwon, D. Ts'o, P. Soliz, B. Zimmerman, J. Pokorny, and R. Kardon, Invest. Ophthalmol. Visual Sci. 47, 715 (2006).
[CrossRef]

K. Grieve and A. Roorda, Invest. Ophthalmol. Visual Sci. 49, 713 (2008).
[CrossRef]

G. Hanazono, K. Tsunoda, Y. Kazato, K. Tsubota, and M. Tanifuji, Invest. Ophthalmol. Visual Sci. 49, 4655 (2008).
[CrossRef]

J. Biomed. Opt. (1)

D. X. Hammer, R. D. Ferguson, T. E. Ustun, C. E. Bigelow, N. V. Iftimia, and R. H. Webb, J. Biomed. Opt. 11, 041126 (2006).
[CrossRef] [PubMed]

Jpn. J. Ophthalmol. (1)

X. C. Yao, Jpn. J. Ophthalmol. 53, 327 (2009).
[CrossRef] [PubMed]

Nature (1)

M. Kahlert, D. R. Pepperberg, and K. P. Hofmann, Nature 345, 537 (1990).
[CrossRef] [PubMed]

Neuroimage (1)

J. L. Schei, M. D. McCluskey, A. J. Foust, X. C. Yao, and D. M. Rector, Neuroimage 40, 1034 (2008).
[CrossRef] [PubMed]

Ophthalmic Surg. Lasers Imaging (1)

D. A. Nelson, S. Krupsky, A. Pollack, E. Aloni, M. Belkin, I. Vanzetta, M. Rosner, and A. Grinvald, Ophthalmic Surg. Lasers Imaging 36, 57 (2005).
[PubMed]

Opt. Express (4)

Opt. Lett. (2)

Proc. Natl. Acad. Sci. USA (2)

D. R. Pepperberg, M. Kahlert, A. Krause, and K. P. Hofmann, Proc. Natl. Acad. Sci. USA 85, 5531 (1988).
[CrossRef] [PubMed]

K. Bizheva, R. Pflug, B. Hermann, B. Povazay, H. Sattmann, P. Qiu, E. Anger, H. Reitsamer, S. Popov, J. R. Taylor, A. Unterhuber, P. Ahnelt, and W. Drexler, Proc. Natl. Acad. Sci. USA 103, 5066 (2006).
[CrossRef] [PubMed]

Visual Neurosci. (1)

Q. V. Hoang, R. A. Linsenmeier, C. K. Chung, and C. A. Curcio, Visual Neurosci. 19, 395 (2002).
[CrossRef]

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Figures (3)

Fig. 1
Fig. 1

(a) Schematic diagram of line-scan confocal system: CO, collimator; CL, cylindrical lens; BS, beam splitter; AOD, acousto-optic deflector; DM, dichroic mirror; L x , spherical lenses. Focal length of the CL is 50 mm. Focal lengths of lenses L1, L2, L3, L4, and L5 are 80, 60, 120, 60, and 100 mm, respectively. (b) Confocal image showing clear cellular structure of individual photoreceptors. In the x direction (i.e., parallel to the focused scanning line), theoretical resolution of the imager is 1.9 μ m ( 0.61 λ /NA). In the y direction (i.e., perpendicular to the focused scanning line), theoretical resolution of the imager is 1.3 μ m ( 0.4 λ / NA ) . (c) Axial resolution of line-scan confocal imaging system was tested to be 12 μ m .

Fig. 2
Fig. 2

Frame-by-frame imaging of IOSs. (a) Raw image of photoreceptors. (b) Prestimulus IOS image. (c) Poststim ulus IOSs image. The raw confocal images were acquired at the speed of 100 frames/s, with NIR imaging light focused at photoreceptor layer. Both prestimulus and poststimulus IOS images were an average over 250 ms. (d) Tracings 1–6 revealed the IOS variation property of local areas pointed by arrowheads 1–6 in (c). Tracing 7 represents integral IOSs by averaging all the pixels of each IOS image. Vertical line indicates the onset of the stimulus. (e) The percentage statistics of activated retinal areas with positive ( > 5 % Δ I / I ) and negative ( < 5 % Δ I / I ) IOSs. The threshold ( 5 % Δ I / I ) was used to reduce the effect of background noise on the statistics. Traces 1 (positive IOSs) and 2 (negative IOSs) are statistic results of experiment trial with stimulus delivered. Traces 3 (positive IOSs) and 4 (negative IOSs) are statistic results of control trial without stimulus.

Fig. 3
Fig. 3

Line-by-line imaging of IOSs: (a) line-by-line raw image, (b) line-by-line IOS image, (c) line-by-line DIOS image. (d) Representative localized IOSs (arrows 1–4). S, visible light stimulus. (e) The statistics of activated retinal area ratios with positive (black) and negative (gray) IOSs. The inset panel shows enlarged picture of early optical response marked by the gray bar. Rapid IOSs occurred within 5 ms after stimulus onset (inset panel). (f) Representative localized DIOSs (arrows 1–4).

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