Abstract

We demonstrate enhanced axial confinement in a temporal focusing setup with a shaped spectrum and a narrow emission filter, achieving a reduction of 1 order of magnitude of the out-of-focus background when compared with conventional point-scanning two-photon microscopy. By rejecting the background in the optical domain, our technique circumvents the noise problems common in other background subtraction techniques.

© 2009 Optical Society of America

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  1. W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
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  2. A. Leray and J. Mertz, Opt. Express 14, 10565 (2006).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef]
  7. D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.
  8. S. Lindek and E. H. K. Stelzer, Opt. Lett. 24, 1505 (1999).
    [CrossRef]
  9. L. Moreaux, O. Sandre, and J. Mertz, J. Opt. Soc. Am. B 17, 1685 (2000).
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    [CrossRef]
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    [CrossRef]
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2009 (1)

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

2008 (3)

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

M. E. Durst, G. Zhu, and C. Xu, Opt. Commun. 281, 1805 (2008).
[CrossRef]

2006 (1)

2005 (5)

2003 (2)

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
[CrossRef]

2001 (1)

J. Mertz and L. Moreaux, Opt. Commun. 196, 330 (2001).
[CrossRef]

2000 (2)

1999 (1)

Cambaliza, M. O.

M. O. Cambaliza and C. Saloma, Opt. Commun. 184, 35 (2000).
[CrossRef]

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

Chu, S.-W.

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

Durst, M.

Durst, M. E.

M. E. Durst, G. Zhu, and C. Xu, Opt. Commun. 281, 1805 (2008).
[CrossRef]

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

Kobat, D.

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

Leray, A.

Lin, C.-H.

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

Lindek, S.

Liu, T.-M.

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

Mertz, J.

Moreaux, L.

Nikitin, A. Y.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

Oron, D.

Saloma, C.

M. O. Cambaliza and C. Saloma, Opt. Commun. 184, 35 (2000).
[CrossRef]

Sandre, O.

Shank, C. V.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

Shroff, H.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

Silberberg, Y.

Stelzer, E. H. K.

Sun, C.-K.

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

Tai, S.-P.

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

Tal, E.

Tang, J.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

van Howe, J.

Vaziri, A.

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

Webb, W. W.

R. M. Williams, W. R. Zipfel, and W. W. Webb, Biophys. J. 88, 1377 (2005).
[CrossRef]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
[CrossRef]

Williams, R. M.

R. M. Williams, W. R. Zipfel, and W. W. Webb, Biophys. J. 88, 1377 (2005).
[CrossRef]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
[CrossRef]

Xu, C.

M. E. Durst, G. Zhu, and C. Xu, Opt. Commun. 281, 1805 (2008).
[CrossRef]

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

G. Zhu, J. van Howe, M. Durst, W. Zipfel, and C. Xu, Opt. Express 13, 2153 (2005).
[CrossRef] [PubMed]

Zhu, G.

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

M. E. Durst, G. Zhu, and C. Xu, Opt. Commun. 281, 1805 (2008).
[CrossRef]

G. Zhu, J. van Howe, M. Durst, W. Zipfel, and C. Xu, Opt. Express 13, 2153 (2005).
[CrossRef] [PubMed]

Zipfel, W.

Zipfel, W. R.

R. M. Williams, W. R. Zipfel, and W. W. Webb, Biophys. J. 88, 1377 (2005).
[CrossRef]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
[CrossRef]

Biophys. J. (1)

R. M. Williams, W. R. Zipfel, and W. W. Webb, Biophys. J. 88, 1377 (2005).
[CrossRef]

J. Biomed. Opt. (1)

S.-W. Chu, S.-P. Tai, T.-M. Liu, C.-K. Sun, and C.-H. Lin, J. Biomed. Opt. 14, 3 (2009).
[CrossRef]

J. Opt. Soc. Am. B (2)

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1368 (2003).
[CrossRef]

Opt. Commun. (3)

M. O. Cambaliza and C. Saloma, Opt. Commun. 184, 35 (2000).
[CrossRef]

M. E. Durst, G. Zhu, and C. Xu, Opt. Commun. 281, 1805 (2008).
[CrossRef]

J. Mertz and L. Moreaux, Opt. Commun. 196, 330 (2001).
[CrossRef]

Opt. Express (4)

Opt. Lett. (1)

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

A. Vaziri, J. Tang, H. Shroff, and C. V. Shank, Proc. Natl. Acad. Sci. USA 105, 20221 (2008).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef] [PubMed]

Other (1)

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

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Figures (4)

Fig. 1
Fig. 1

Experimental SFG SSTF setup.

Fig. 2
Fig. 2

Measured spectrum as a function of axial position. Inset, desired filter bandwidth to detect only SFG contribution of the signal.

Fig. 3
Fig. 3

Measured thin film axial scans in the (a) wide-field and (b) line-scanning SSTF configurations, with corresponding (a) line and (b) point SHG scans (log scale).

Fig. 4
Fig. 4

Measured axial response of two collagen films placed 45 μ m apart with (solid curve) and without (dashed curve) spectral shaping (log scale).

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