Abstract

We demonstrate a temporally decorrelated, multifocal multiphoton microscope. Using an etalon, we split the 800-nm light from either an ultrashort-pulsed Ti:Al2O3 oscillator or a Ti:Al2O3 regenerative amplifier into an array of beamlets that are delayed with respect to one another in time. The collimated beams overlap at slightly different input angles at the entrance pupil of a 1.25-numerical aperture oil-immersion objective to produce an array of foci that are temporally decorrelated at the focal plane of the objective. The temporal decorrelation eliminates any interference among the foci and permits multifocal multiphoton imaging with the resolution of single-point illumination.

© 2000 Optical Society of America

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References

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1998

J. Bewersdorf, R. Pick, and S. W. Hell, Opt. Lett. 23, 655 (1998).
[CrossRef]

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

1997

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

S. Mauro, O. Nakamura, and S. Kawata, Opt. Lett. 22, 132 (1997).
[CrossRef]

1996

1990

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

Barad, Y.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Bewersdorf, J.

Brakenhoff, G. J.

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

Buist, A. H.

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

Denk, W.

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

Eisenberg, H.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Hell, S. W.

Horowitz, M.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Juskaitis, R.

Kawata, S.

Kozubek, M.

Mauro, S.

Muller, M.

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

Nakamura, O.

Neil, M. A. A.

Pick, R.

Silberberg, Y.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

Squier, J.

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

Strickler, J. H.

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

Webb, W. W.

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

Wilson, T.

Appl. Phys. Lett.

Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg, Appl. Phys. Lett. 70, 922 (1997).
[CrossRef]

J. Microsc.

A. H. Buist, M. Muller, J. Squier, and G. J. Brakenhoff, J. Microsc. 192, 217 (1998).
[CrossRef]

Opt. Lett.

Science

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

Temporally decorrelated multiphoton microscope. An etalon divides the beam into beamlets, which are then manipulated to produce temporally decorrelated foci in the sample: DM, dichroic mirror; m1, m2 scan mirrors.

Fig. 2
Fig. 2

Line of foci produced by the time-decorrelated multifocal microscope in a cell containing Coumerin dye.

Fig. 3
Fig. 3

Measurement of the PSF for eight foci produced with a Ti:sapphire oscillator and comparison with theory. The theory assumes a 1.25-N.A. Gaussian beam focus.

Fig. 4
Fig. 4

Image of a 15µm polystyrene fluorescent microsphere taken with a Ti:sapphire oscillator and a time-decorrelated multifocal microscope. The foci were asynchronously rastered with the scanning mirrors in the microscope, and the integration time was 60 s.

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