Abstract

We demonstrate selective two-photon excited fluorescence microscopy with shaped pulses produced with a simple yet efficient scheme based on dispersive optical components. The pulse train from a broadband oscillator is split into two subtrains that are sent through different amounts of glass. Beam recombination results in pulse-shape switching at a rate of 150MHz. Time-resolved photon counting detection then provides two simultaneous images resulting from selective two-photon excitation, as demonstrated in a live embryo. Although less versatile than programmable pulse-shaping devices, this novel arrangement significantly improves the performance of selective microscopy using broadband shaped pulses while simplifying the experimental setup.

© 2010 Optical Society of America

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2007 (1)

2006 (2)

2004 (3)

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

M. Comstock, V. Lozovoy, I. Pastirk, and M. Dantus, Opt. Express 12, 1061 (2004).
[PubMed]

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

2003 (2)

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

1998 (1)

D. Meshulach and Y. Silberberg, Nature 396, 239 (1998).

1984 (1)

Amir, W.

Barzda, V.

Beaurepaire, E.

Borri, P.

I. Rocha-Mendoza, W. Langbein, P. Watson, and P. Borri, Opt. Lett. 34, 2258 (2009).
[PubMed]

W. Langbein, I. Rocha-Mendoza, and P. Borri, Appl. Phys. Lett. 95, 081109 (2009).

Boudoux, C.

Carriles, R.

Chandler, E.

Comstock, M.

M. Comstock, V. Lozovoy, I. Pastirk, and M. Dantus, Opt. Express 12, 1061 (2004).
[PubMed]

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

Cruz, J. D.

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

Dantus, M.

V. Lozovoy, B. Xu, J. Shane, and M. Dantus, Phys. Rev. A 74, 041805 (2006).

M. Comstock, V. Lozovoy, I. Pastirk, and M. Dantus, Opt. Express 12, 1061 (2004).
[PubMed]

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

Debarre, D.

Ding, S.

Durfee, C.

Farge, E.

Field, J.

Fork, R.

Gordon, J.

Hoover, E.

Isobe, K.

Joffre, M.

Kannari, F.

Kawano, H.

Labroille, G.

Langbein, W.

I. Rocha-Mendoza, W. Langbein, P. Watson, and P. Borri, Opt. Lett. 34, 2258 (2009).
[PubMed]

W. Langbein, I. Rocha-Mendoza, and P. Borri, Appl. Phys. Lett. 95, 081109 (2009).

Lozovoy, V.

V. Lozovoy, B. Xu, J. Shane, and M. Dantus, Phys. Rev. A 74, 041805 (2006).

M. Comstock, V. Lozovoy, I. Pastirk, and M. Dantus, Opt. Express 12, 1061 (2004).
[PubMed]

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

Martin, J.-L.

Martinez, O.

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D. Meshulach and Y. Silberberg, Nature 396, 239 (1998).

Midorikawa, K.

Miyawaki, A.

Mizuno, H.

Ogilvie, J. P.

Olivier, N.

Pastirk, I.

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

M. Comstock, V. Lozovoy, I. Pastirk, and M. Dantus, Opt. Express 12, 1061 (2004).
[PubMed]

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

Pillai, R. S.

Planchon, T.

Rocha-Mendoza, I.

W. Langbein, I. Rocha-Mendoza, and P. Borri, Appl. Phys. Lett. 95, 081109 (2009).

I. Rocha-Mendoza, W. Langbein, P. Watson, and P. Borri, Opt. Lett. 34, 2258 (2009).
[PubMed]

Shane, J.

V. Lozovoy, B. Xu, J. Shane, and M. Dantus, Phys. Rev. A 74, 041805 (2006).

Sheetz, K.

Silberberg, Y.

D. Meshulach and Y. Silberberg, Nature 396, 239 (1998).

Solinas, X.

Squier, J.

Suda, A.

Tanaka, M.

Veilleux, I.

Walowicz, K.

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

I. Pastirk, J. D. Cruz, K. Walowicz, V. Lozovoy, and M. Dantus, Opt. Express 11, 1695 (2003).
[PubMed]

Watson, P.

Xu, B.

V. Lozovoy, B. Xu, J. Shane, and M. Dantus, Phys. Rev. A 74, 041805 (2006).

Appl. Opt. (1)

Appl. Phys. Lett. (1)

W. Langbein, I. Rocha-Mendoza, and P. Borri, Appl. Phys. Lett. 95, 081109 (2009).

J. Chem. Phys. (1)

V. Lozovoy, I. Pastirk, K. Walowicz, and M. Dantus, J. Chem. Phys. 118, 3187 (2003).

J. Phys. Chem. A (1)

J. D. Cruz, I. Pastirk, V. Lozovoy, K. Walowicz, and M. Dantus, J. Phys. Chem. A 108, 53 (2004).

Nature (1)

D. Meshulach and Y. Silberberg, Nature 396, 239 (1998).

Opt. Express (6)

Opt. Lett. (3)

Phys. Rev. A (1)

V. Lozovoy, B. Xu, J. Shane, and M. Dantus, Phys. Rev. A 74, 041805 (2006).

Proc. Natl. Acad. Sci. U.S.A. (1)

J. D. Cruz, I. Pastirk, M. Comstock, V. Lozovoy, and M. Dantus, Proc. Natl. Acad. Sci. U.S.A. 101, 16996 (2004).

Supplementary Material (1)

» Media 1: AVI (10974 KB)     

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Figures (3)

Fig. 1
Fig. 1

Schematic plot of the group delay introduced by (a) the prism compressor, (b) the optics of the microscope, and (c) the optics of the microscope plus an additional dispersive glass placed in the beam path, and total group delay without ( a + b ) and with ( a + c ) the dispersive glass. The additional dispersion results in a shift of the extremum (diamond).

Fig. 2
Fig. 2

Experimental setup: HWP, half-wave plate; PBS, polarizing beam splitter; GM, galvanometer-mounted mirrors; T rep , laser period ( 1 / T rep 75 MHz ); c, speed of light; c T rep 4 m .

Fig. 3
Fig. 3

Imaging with shaped pulses. (a), (b) Drosophila images obtained with blueshifted pulses preferentially exciting (a) blue endogenous fluorescence and (b) redshifted pulses preferentially exciting GFP. (c) Corresponding two-photon spectra measured with a BBO crystal for the blueshifted and redshifted pulses. (d) Combination of (a) and (b) according to the color scale. Refer to movie file (Media 1).

Equations (4)

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φ 1 ( ω ) = 1 6 φ ( ω ω 1 ) 3 ,
τ 1 ( ω ) = d φ 1 d ω = 1 2 φ ( ω ω 1 ) 2 .
τ 2 ( ω ) = τ 1 ( ω ) + φ glass ( ω ω 0 ) ,
= 1 2 φ ( ω ω 1 + φ glass φ ) 2 + Cst .

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