Abstract

We introduce a direct wavefront sensing method using structures labeled with fluorescent proteins in tissues as guide stars. An adaptive optics confocal microscope using this method is demonstrated for imaging of mouse brain tissue. A dendrite and a cell body of a neuron labeled with yellow fluorescent protein are tested as guide stars without injection of other fluorescent labels. Photobleaching effects are also analyzed. The results shows increased image contrast and 3× improvement in the signal intensity for fixed mouse tissues at depths of 70μm.

© 2011 Optical Society of America

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2011 (3)

2010 (2)

O. Azucena, J. Crest, J. Cao, W. Sullivan, P. Kner, D. Gavel, D. Dillon, S. Olivier, and J. Kubby, Opt. Express 18, 17521 (2010).
[CrossRef] [PubMed]

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

2009 (1)

2007 (2)

2006 (2)

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

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

2002 (2)

1999 (1)

Azucena, O.

Beverage, J.

J. Beverage, R. Shack, and M. Descour, J. Microsc. 205, 61 (2002).
[CrossRef] [PubMed]

Bifano, T. G.

Biss, D. P.

Booth, M. J.

Botcherby, E. J.

Brase, J. M.

Burns, S. A.

Cao, J.

Chen, D.

Côté, D.

Crest, J.

Dainty, J. C.

Debarre, D.

Denk, W.

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

Descour, M.

J. Beverage, R. Shack, and M. Descour, J. Microsc. 205, 61 (2002).
[CrossRef] [PubMed]

Diaz Santana Haro, L.

Dillon, D.

O. Azucena, J. Crest, J. Cao, W. Sullivan, P. Kner, D. Gavel, D. Dillon, S. Olivier, and J. Kubby, Opt. Express 18, 17521 (2010).
[CrossRef] [PubMed]

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Fernandez, B.

Fu, M.

Fusco, T.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Garcia, D.

Gavel, D.

Gavel, D. T.

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

L. A. Poyneer, D. T. Gavel, and J. M. Brase, J. Opt. Soc. Am. A 19, 2100 (2002).
[CrossRef]

Grigsby, B.

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Johnson, L. C.

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Kner, P.

Kotadia, S.

O. Azucena, J. Crest, S. Kotadia, W. Sullivan, X. Tao, M. Reinig, D. Gavel, S. Olivier, and J. Kubby, Opt. Lett. 36, 825 (2011).
[CrossRef] [PubMed]

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, Proc. SPIE 7931, 79310J (2011).
[CrossRef]

Kubby, J.

Lakowicz, J. R.

J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, 2006).
[CrossRef]

Lin, C. P.

Macintosh, B. A.

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Mack-Bucher, J. A.

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

Michau, V.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Morzinski, K.

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Nicolle, M.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Olivier, S.

Poyneer, L. A.

Reinig, M.

O. Azucena, J. Crest, S. Kotadia, W. Sullivan, X. Tao, M. Reinig, D. Gavel, S. Olivier, and J. Kubby, Opt. Lett. 36, 825 (2011).
[CrossRef] [PubMed]

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, Proc. SPIE 7931, 79310J (2011).
[CrossRef]

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Rousset, G.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Rueckel, M.

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

Shack, R.

J. Beverage, R. Shack, and M. Descour, J. Microsc. 205, 61 (2002).
[CrossRef] [PubMed]

Srinivas, S.

Sullivan, W.

Sumorok, D.

Tao, X.

Thomas, S.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Tokovinin, A.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Veilleux, I.

Watanabe, T.

Webb, R. H.

Wilson, T.

Zamiri, P.

Zhou, Y.

Zuo, Y.

J. Microsc. (1)

J. Beverage, R. Shack, and M. Descour, J. Microsc. 205, 61 (2002).
[CrossRef] [PubMed]

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

Mon. Not. R. Astron. Soc. (1)

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, Mon. Not. R. Astron. Soc. 371, 323 (2006).
[CrossRef]

Opt. Express (1)

Opt. Lett. (5)

Phil. Trans. R. Soc. A (1)

M. J. Booth, Phil. Trans. R. Soc. A 365, 2829 (2007).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. USA (1)

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

Proc. SPIE (2)

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, Proc. SPIE 7931, 79310J (2011).
[CrossRef]

K. Morzinski, L. C. Johnson, D. T. Gavel, B. Grigsby, D. Dillon, M. Reinig, and B. A. Macintosh, Proc. SPIE 7736, 77361O (2010).
[CrossRef]

Other (1)

J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, 2006).
[CrossRef]

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Figures (4)

Fig. 1
Fig. 1

AO confocal microscope setup. DB, dichroic BS; I1 and I2, irises; F2, excitation filter; F1 and F3, emission filters; BB, beam blocker. See text for details.

Fig. 2
Fig. 2

Wavefront measurements using YFP- labeled structures as guide stars. The excitation light source is focused on a cell body of a neuron in (a) and on a dendrite in (b). The scale bars are 5 μm . The crossed lines indicate the location of the focus point. The wavefront measurements are shown for a dendrite in (c) and a cell body in (d) (x, y scale in subapertures).

Fig. 3
Fig. 3

Photobleaching analysis. Normalized intensity at the focal point on the dendrite.

Fig. 4
Fig. 4

Confocal fluorescence imaging of mouse brain tissue. The maximum intensity projection image (a) before and (b) after correction. The dashed boxes indicate enlarged images (c) before and (d) after correction. (e) Wavefront error after correction (x, y scale in subapertures). (f) Intensity profiles along the dashed lines in (c) and (d). The scale bar is 5 μm .

Equations (1)

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d diffraction_limit = 2.44 λ 2 NA D 0 d l ,

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