Abstract

We have demonstrated successful two-photon excitation fluorescence bioimaging using a high-power pulsed all-semiconductor laser. Toward this purpose, we developed a pulsed light source consisting of a mode-locked laser diode and a two-stage diode laser amplifier. This pulsed light source provided optical pulses of 5ps duration and having a maximum peak power of over 100 W at a wavelength of 800nm and a repetition frequency of 500MHz.

© 2007 Optical Society of America

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References

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2007 (1)

2006 (1)

2005 (1)

2003 (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1369 (2003).
[CrossRef] [PubMed]

2001 (1)

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, Biophys. J. 80, 1568 (2001).
[CrossRef] [PubMed]

1999 (1)

A. Zumbush, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

1990 (1)

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

1989 (1)

G. P. Agrawal and N. A. Olsson, IEEE J. Quantum Electron. 25, 2297 (1989).
[CrossRef]

Biophys. J. (1)

L. Moreaux, O. Sandre, S. Charpak, M. Blanchard-Desce, and J. Mertz, Biophys. J. 80, 1568 (2001).
[CrossRef] [PubMed]

IEEE J. Quantum Electron. (1)

G. P. Agrawal and N. A. Olsson, IEEE J. Quantum Electron. 25, 2297 (1989).
[CrossRef]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1369 (2003).
[CrossRef] [PubMed]

Opt. Express (3)

Phys. Rev. Lett. (1)

A. Zumbush, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
[CrossRef]

Science (1)

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

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Figures (4)

Fig. 1
Fig. 1

Schematic configuration of the all semiconductor laser picosecond light pulse source. MLLD, mode-locked laser diode; BPF, bandpass filter; OI, optical isolator; SOA, semiconductor optical amplifier; lens, collimating/focusing lens.

Fig. 2
Fig. 2

(a) SHG autocorrelation trace of the amplified optical pulses showing a pulse width of 5.2 ps ( sech 2 pulse shape assumed). (b) Optical spectra of the mode-locked laser pulse and the amplified pulses showing a wavelength of 783 nm .

Fig. 3
Fig. 3

Photograph indicating bright two-photon excited fluorescence in a cell containing Coumarin 480 dye excited by 20 W peak-power light pulses. The peak optical power density of 1 MW cm 2 at the cell can be estimated by considering the optical systems with a 20 × expansion and near-field pattern of LD system.

Fig. 4
Fig. 4

Two-photon excited fluorescence image of glomeruli and convoluted tubules in mouse kidney tissues stained with Alexa Fluor 488. The incident average optical power was 98 mW ( 32 W peak power).

Equations (1)

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I = k T τ P a v 2 .

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