Abstract

We demonstrate image-based aberration correction in a third-harmonic generation (THG) microscope. We describe a robust, mostly sample-independent correction scheme relying on prior measurement of the influence of aberration modes produced by a deformable mirror on the quality of THG images. We find that using image sharpness as an image quality metric, correction of N aberration modes is achieved using 2(2N+1) measurements in a variety of samples. We also report aberration correction in combined multiharmonic and two-photon excited fluorescence experiments. Finally, we demonstrate time-dependent adaptive THG imaging in developing embryonic tissue.

© 2009 Optical Society of America

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2009 (1)

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

2008 (2)

2006 (1)

M. Rueckel, J. Mack-Bucher, and W. Denk, Proc. Natl. Acad. Sci. USA 103, 17137 (2006).
[CrossRef] [PubMed]

2005 (1)

2004 (1)

2003 (1)

2002 (1)

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

1997 (1)

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

1985 (1)

Barad, Y.

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

Beaurepaire, E.

Booth, M. J.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

D. Débarre, E. J. Botcherby, M. J. Booth, and T. Wilson, Opt. Express 16, 9290 (2008).
[CrossRef] [PubMed]

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

Botcherby, E. J.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

D. Débarre, E. J. Botcherby, M. J. Booth, and T. Wilson, Opt. Express 16, 9290 (2008).
[CrossRef] [PubMed]

Burns, D.

Débarre, D.

Denk, W.

M. Rueckel, J. Mack-Bucher, and W. Denk, Proc. Natl. Acad. Sci. USA 103, 17137 (2006).
[CrossRef] [PubMed]

Eisenberg, H.

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

Farge, E.

Girkin, J. M.

Herloski, R.

Horowitz, M.

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

Juškaitis, R.

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

Mack-Bucher, J.

M. Rueckel, J. Mack-Bucher, and W. Denk, Proc. Natl. Acad. Sci. USA 103, 17137 (2006).
[CrossRef] [PubMed]

Marsh, P. N.

Moulia, B.

Neil, M. A. A.

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

Olivier, N.

Rueckel, M.

M. Rueckel, J. Mack-Bucher, and W. Denk, Proc. Natl. Acad. Sci. USA 103, 17137 (2006).
[CrossRef] [PubMed]

Schanne-Klein, M.-C.

Silberberg, Y.

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

Srinivas, S.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

Supatto, W.

Watanabe, T.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

Wilson, T.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, Opt. Lett. 16, 2495 (2009).
[CrossRef]

D. Débarre, E. J. Botcherby, M. J. Booth, and T. Wilson, Opt. Express 16, 9290 (2008).
[CrossRef] [PubMed]

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

Appl. Phys. Lett. (1)

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

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

Opt. Express (3)

Opt. Lett. (3)

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

M. Rueckel, J. Mack-Bucher, and W. Denk, Proc. Natl. Acad. Sci. USA 103, 17137 (2006).
[CrossRef] [PubMed]

M. J. Booth, M. A. A. Neil, R. Juškaitis, and T. Wilson, Proc. Natl. Acad. Sci. USA 99, 5788 (2002).
[CrossRef] [PubMed]

Supplementary Material (4)

» Media 1: AVI (13564 KB)     
» Media 2: AVI (4159 KB)     
» Media 3: AVI (9838 KB)     
» Media 4: AVI (3950 KB)     

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Figures (4)

Fig. 1
Fig. 1

(a) Experimental setup: λ 4 , quarter wave plate; PBS, polarizing beamsplitter; DBS, dichroic beamsplitter. Solid lines, excitation path; gray, DM characterization path (not used during AC); dotted lines, emission path; dashed lines, conjugated Fourier planes. (b) Modes X i used for correction, as measured on the SH sensor. The phase is color coded from purple to red (online) with a brightness proportional to excitation intensity. The last image (lower right) shows the intensity profile alone.

Fig. 2
Fig. 2

(a) THG images of an elderberry stem slice (left) before and (right) after AC on the middle plane of the z stack. Scale bars, 20 μ m . (b) Profiles along the lines in (a). (c) Averaged influence matrix A. (d) TH image brightness as a function of the number of AC runs, using A (squares) or assuming independent modes (circles). (e) Corresponding final phases (in radians). The top phase was applied for the corrected image in (a).

Fig. 3
Fig. 3

(a) (Media 1, high resolution; Media 2, low resolution) Combined 2PEF (online)/THG (interior) images of coverslip-covered lily pollen grains, without AC (left), with THG (center), and 2PEF (right) AC. Scale bar, 50 μ m . (b) Metric versus aberration amplitude in mode 1 [see Fig. 1b] for THG sharpness (bottom curve), 2PEF brightness (top curve) and THG brightness (middle curve). Lines, Gaussian fits. (c) Applied phase and 2PEF (left block)/THG (right block) signal increase in (a): left, THG AC; right, 2PEF AC. (d) Combined THG (light)/SHG (dark) images of a coverslip-covered lily anther slice, before (left), and after (right) THG AC. (e) Corresponding AC phase and signal increase. Scale bar, 20 μ m .

Fig. 4
Fig. 4

(a) (Media 3, high resolution; Media 4, low resolutions) Drosophila development imaging without (top) and with (bottom) time-resolved THG-AC. Inset, correction phase (in radians). (b) Zoom on the square in (a). (c) Signal increase after AC. (d) AC amplitude in three modes and total amplitude.

Equations (4)

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I X i X j d r I d r I X i d r I X j d r ( I d r ) 2 = δ i j ,
M M 0 exp [ i , j α i j a i a j ] = M 0 exp [ a T A a ] ,
a = { b i 2 ln M i + ln M i 2 ln M 0 ln M i ln M i + } .
a = A 1 { α i i b i 2 ln M i + ln M i 2 ln M 0 ln M i ln M i + } .

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