Abstract

We developed a novel addressable discrete-line-scanning multiphoton microscope with high lateral and axial resolutions based on a spatial light modulator. Our discrete-line-focus design eliminates the cross talk that occurs in conventional one-dimensional line-scanning multiphoton microscopies. Additionally, a phase-only spatial light modulator is able to scan only a sample’s target area by generating a specific discrete line focus according to the shape and location of the target area. Compared with other multiphoton microscopies, this technique shortens scanning time and minimizes photodamage by concentrating scanning energy and dwell time on the area of interest.

© 2012 Optical Society of America

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References

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2008 (1)

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

2003 (1)

2001 (2)

S. W. Hell and V. Andresen, J. Microsc. 202, 457 (2001).
[CrossRef]

S. W. Hell, V. Andresen, and A. Egner, Opt. Lett. 26, 75 (2001).
[CrossRef]

2000 (1)

1998 (1)

1996 (1)

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

1990 (1)

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

1972 (1)

R. W. Gerchber and W. O. Saxton, Optik 35, 237 (1972).

1968 (1)

Andresen, V.

S. W. Hell, V. Andresen, and A. Egner, Opt. Lett. 26, 75 (2001).
[CrossRef]

S. W. Hell and V. Andresen, J. Microsc. 202, 457 (2001).
[CrossRef]

Antolini, R.

Araya, R.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Athey, B.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Bewersdorf, J.

Bliton, A. C.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Brakenhoff, G. J.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Choudhury, A.

Denk, W.

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

Egger, M. D.

Egner, A.

Froner, E.

Galambos, R.

Gerchber, R. W.

R. W. Gerchber and W. O. Saxton, Optik 35, 237 (1972).

Hadravsk, M.

Hell, S. W.

Nikolenko, V.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Norris, T.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Pavone, F. S.

Peterka, D. S.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Petran, M.

Pick, R.

Sacconi, L.

Saxton, W. O.

R. W. Gerchber and W. O. Saxton, Optik 35, 237 (1972).

Squier, J.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Strickler, J. H.

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

Taghizadeh, M. R.

Wade, M. H.

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

Watson, B. O.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Webb, W. W.

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

Woodruff, A.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Yuste, R.

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

Front Neural Circuits (1)

V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front Neural Circuits 2, 5 (2008).

J. Microsc. (2)

G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
[CrossRef]

S. W. Hell and V. Andresen, J. Microsc. 202, 457 (2001).
[CrossRef]

J. Opt. Soc. Am. (1)

J. Opt. Soc. Am. A (1)

Opt. Lett. (3)

Optik (1)

R. W. Gerchber and W. O. Saxton, Optik 35, 237 (1972).

Science (1)

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

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Figures (3)

Fig. 1.
Fig. 1.

Schematic optical setup of the ADLS-MMM. PBS, polarized beam splitter; GS, 2D galvo scanner; SLM, spatial light modulator; zero-order beam blocker, a small piece of black-painted aluminum foil on a glass slide used to block the unwanted zero-order bright spot; L1, L2, beam expander (f=30 and 100 mm, respectively); L3< scanning lens (f=200mm); L4, tube lens (f=180mm); Objective, water-immersion objective, 60×, NA=1.0; M1, M2, M3, M4, mirrors.

Fig. 2.
Fig. 2.

Design of the discrete lines with (a) shorter distance, (b) longer distance between adjacent points, and (c) image of the line.

Fig. 3.
Fig. 3.

(a) Two-photon image of adult rat cardiomyocyte in full FOV obtained by ADLS-MMM, (b) axial-stack two-photon images of the cell.

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