Abstract

Spatially and temporally dependent optical aberrations induced by the inhomogeneous refractive index of live samples limit the resolution of live dynamic imaging. We introduce an adaptive optical microscope with a direct wavefront sensing method using a Shack-Hartmann wavefront sensor and fluorescent protein guide-stars for live imaging. The results of imaging Drosophila embryos demonstrate its ability to correct aberrations and achieve near diffraction limited images of medial sections of large Drosophila embryos. GFP-polo labeled centrosomes can be observed clearly after correction but cannot be observed before correction. Four dimensional time lapse images are achieved with the correction of dynamic aberrations. These studies also demonstrate that the GFP-tagged centrosome proteins, Polo and Cnn, serve as excellent biological guide-stars for adaptive optics based microscopy.

© 2012 OSA

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  1. M. J. Booth, “Adaptive optics in microscopy,” Phil. Trans. R. Soc. A–Math. Phys. Eng. Sci. 365, 2829–2843 (2007).
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  3. J. Porter, H. Queener, J. Lin, K. Thorn, and A. A. S. Awwal, Adaptive Optics for Vision Science: Principles, Practices, Design and Applications, (Wiley, 2006).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  9. N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods 7(2), 141–147 (2010).
    [CrossRef] [PubMed]
  10. D. E. Milkie, E. Betzig, and N. Ji, “Pupil-segmentation-based adaptive optical microscopy with full-pupil illumination,” Opt. Lett. 36(21), 4206–4208 (2011).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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  21. J. W. Hardy, Adaptive Optics for Astronomical Telescopes (Oxford University Press, 1998).
  22. M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [PubMed]
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2012 (1)

2011 (6)

2010 (2)

2009 (2)

2007 (2)

M. J. Booth, “Adaptive optics in microscopy,” Phil. Trans. R. Soc. A–Math. Phys. Eng. Sci. 365, 2829–2843 (2007).

J. Zhang and T. L. Megraw, “Proper recruitment of gamma-tubulin and D-TACC/Msps to embryonic Drosophila centrosomes requires Centrosomin Motif 1,” Mol. Biol. Cell 18(10), 4037–4049 (2007).
[CrossRef] [PubMed]

2006 (2)

M. Rueckel, J. A. Mack-Bucher, and W. Denk, “Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing,” Proc. Natl. Acad. Sci. U.S.A. 103(46), 17137–17142 (2006).
[CrossRef] [PubMed]

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

2004 (2)

L. A. Poyneer and B. A. Macintosh, “Spatially filtered wave-front sensor for high-order adaptive optics,” J. Opt. Soc. Am. A 21(5), 810–819 (2004).
[CrossRef] [PubMed]

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

2003 (1)

2002 (2)

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

L. A. Poyneer, D. T. Gavel, and J. M. Brase, “Fast wave-front reconstruction in large adaptive optics systems with use of the Fourier transform,” J. Opt. Soc. Am. A 19(10), 2100–2111 (2002).
[CrossRef]

1999 (1)

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Amorim, I.

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Andilla, J.

Artigas, D.

Aviles-Espinosa, R.

Azucena, O.

Beaurepaire, E.

Betzig, E.

D. E. Milkie, E. Betzig, and N. Ji, “Pupil-segmentation-based adaptive optical microscopy with full-pupil illumination,” Opt. Lett. 36(21), 4206–4208 (2011).
[CrossRef] [PubMed]

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods 7(2), 141–147 (2010).
[CrossRef] [PubMed]

Booth, M. J.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, “Image-based adaptive optics for two-photon microscopy,” Opt. Lett. 34(16), 2495–2497 (2009).
[CrossRef] [PubMed]

M. J. Booth, “Adaptive optics in microscopy,” Phil. Trans. R. Soc. A–Math. Phys. Eng. Sci. 365, 2829–2843 (2007).

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

Botcherby, E. J.

Brase, J. M.

Burns, D.

Cao, J.

Chen, D. C.

Costa, M.

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Crest, J.

Débarre, D.

Denk, W.

M. Rueckel, J. A. Mack-Bucher, and W. Denk, “Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing,” Proc. Natl. Acad. Sci. U.S.A. 103(46), 17137–17142 (2006).
[CrossRef] [PubMed]

Dillon, D.

Facomprez, A.

Fernandez, B.

Fu, M.

Fusco, T.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Garcia, D.

Gavel, D.

Gavel, D. T.

Girkin, J.

Ji, N.

D. E. Milkie, E. Betzig, and N. Ji, “Pupil-segmentation-based adaptive optical microscopy with full-pupil illumination,” Opt. Lett. 36(21), 4206–4208 (2011).
[CrossRef] [PubMed]

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods 7(2), 141–147 (2010).
[CrossRef] [PubMed]

Juskaitis, R.

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

Kner, P.

Kotadia, S.

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, “Adaptive optics wide-field microscope corrections using a MEMS DM and Shack-Hartmann wavefront sensor,” Proc. SPIE 7931, 79310J (2011).
[CrossRef]

O. Azucena, J. Crest, S. Kotadia, W. Sullivan, X. Tao, M. Reinig, D. Gavel, S. Olivier, and J. Kubby, “Adaptive optics wide-field microscopy using direct wavefront sensing,” Opt. Lett. 36(6), 825–827 (2011).
[CrossRef] [PubMed]

Kubby, J.

Levecq, X.

Loza-Alvarez, P.

Macintosh, B. A.

Mack-Bucher, J. A.

M. Rueckel, J. A. Mack-Bucher, and W. Denk, “Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing,” Proc. Natl. Acad. Sci. U.S.A. 103(46), 17137–17142 (2006).
[CrossRef] [PubMed]

Marsh, P.

Megraw, T. L.

J. Zhang and T. L. Megraw, “Proper recruitment of gamma-tubulin and D-TACC/Msps to embryonic Drosophila centrosomes requires Centrosomin Motif 1,” Mol. Biol. Cell 18(10), 4037–4049 (2007).
[CrossRef] [PubMed]

Michau, V.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Milkie, D. E.

D. E. Milkie, E. Betzig, and N. Ji, “Pupil-segmentation-based adaptive optical microscopy with full-pupil illumination,” Opt. Lett. 36(21), 4206–4208 (2011).
[CrossRef] [PubMed]

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods 7(2), 141–147 (2010).
[CrossRef] [PubMed]

Moutinho-Santos, T.

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Neil, M. A.

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

Nicolle, M.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Nieto, M.

Olarte, O. E.

Olivier, N.

Olivier, S.

Porcar-Guezenec, R.

Poyneer, L. A.

Reinig, M.

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, “Adaptive optics wide-field microscope corrections using a MEMS DM and Shack-Hartmann wavefront sensor,” Proc. SPIE 7931, 79310J (2011).
[CrossRef]

O. Azucena, J. Crest, S. Kotadia, W. Sullivan, X. Tao, M. Reinig, D. Gavel, S. Olivier, and J. Kubby, “Adaptive optics wide-field microscopy using direct wavefront sensing,” Opt. Lett. 36(6), 825–827 (2011).
[CrossRef] [PubMed]

Rousset, G.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Rueckel, M.

M. Rueckel, J. A. Mack-Bucher, and W. Denk, “Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing,” Proc. Natl. Acad. Sci. U.S.A. 103(46), 17137–17142 (2006).
[CrossRef] [PubMed]

Sampaio, P.

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Schwertner, M.

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

Srinivas, S.

Sullivan, W.

Sunkel, C. E.

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Tao, X.

Thomas, S.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Tokovinin, A.

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Watanabe, T.

Wilson, T.

D. Débarre, E. J. Botcherby, T. Watanabe, S. Srinivas, M. J. Booth, and T. Wilson, “Image-based adaptive optics for two-photon microscopy,” Opt. Lett. 34(16), 2495–2497 (2009).
[CrossRef] [PubMed]

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

Zhang, J.

J. Zhang and T. L. Megraw, “Proper recruitment of gamma-tubulin and D-TACC/Msps to embryonic Drosophila centrosomes requires Centrosomin Motif 1,” Mol. Biol. Cell 18(10), 4037–4049 (2007).
[CrossRef] [PubMed]

Zuo, Y.

Biol. Cell (1)

T. Moutinho-Santos, P. Sampaio, I. Amorim, M. Costa, and C. E. Sunkel, “In vivo localisation of the mitotic POLO kinase shows a highly dynamic association with the mitotic apparatus during early embryogenesis in Drosophila,” Biol. Cell 91(8), 585–596 (1999).
[PubMed]

Biomed. Opt. Express (1)

J. Microsc. (1)

M. Schwertner, M. J. Booth, M. A. Neil, and T. Wilson, “Measurement of specimen-induced aberrations of biological samples using phase stepping interferometry,” J. Microsc. 213(1), 11–19 (2004).
[CrossRef] [PubMed]

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

Mol. Biol. Cell (1)

J. Zhang and T. L. Megraw, “Proper recruitment of gamma-tubulin and D-TACC/Msps to embryonic Drosophila centrosomes requires Centrosomin Motif 1,” Mol. Biol. Cell 18(10), 4037–4049 (2007).
[CrossRef] [PubMed]

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

S. Thomas, T. Fusco, A. Tokovinin, M. Nicolle, V. Michau, and G. Rousset, “Comparison of centroid computation algorithms in a Shack-Hartmann sensor,” Mon. Not. R. Astron. Soc. 371(1), 323–336 (2006).
[CrossRef]

Nat. Methods (1)

N. Ji, D. E. Milkie, and E. Betzig, “Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues,” Nat. Methods 7(2), 141–147 (2010).
[CrossRef] [PubMed]

Opt. Express (3)

Opt. Lett. (6)

Phil. Trans. R. Soc. A–Math. Phys. Eng. Sci. (1)

M. J. Booth, “Adaptive optics in microscopy,” Phil. Trans. R. Soc. A–Math. Phys. Eng. Sci. 365, 2829–2843 (2007).

Proc. Natl. Acad. Sci. U.S.A. (2)

M. J. Booth, M. A. Neil, R. Juskaitis, and T. Wilson, “Adaptive aberration correction in a confocal microscope,” Proc. Natl. Acad. Sci. U.S.A. 99(9), 5788–5792 (2002).
[CrossRef] [PubMed]

M. Rueckel, J. A. Mack-Bucher, and W. Denk, “Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing,” Proc. Natl. Acad. Sci. U.S.A. 103(46), 17137–17142 (2006).
[CrossRef] [PubMed]

Proc. SPIE (1)

O. Azucena, X. Tao, J. Crest, S. Kotadia, W. Sullivan, D. Gavel, M. Reinig, S. Olivier, and J. Kubby, “Adaptive optics wide-field microscope corrections using a MEMS DM and Shack-Hartmann wavefront sensor,” Proc. SPIE 7931, 79310J (2011).
[CrossRef]

Other (6)

M. Gu, Advanced Optical Imaging Theory (Springer-Verlag, New York, 1999).

M. Gu, Principles of Three-Dimensional Imaging in Confocal Microscopes (World Scientific, Singapore, 1996).

J. W. Hardy, Adaptive Optics for Astronomical Telescopes (Oxford University Press, 1998).

R. K. Tyson, Principles of Adaptive Optics (Academic, 1991).

J. Porter, H. Queener, J. Lin, K. Thorn, and A. A. S. Awwal, Adaptive Optics for Vision Science: Principles, Practices, Design and Applications, (Wiley, 2006).

W. F. Rothwell and W. Sullivan, “Fluorescent analysis of drosophila embryos,” in Drosophila Protocols, W. Sullivan, M. Ashburner and R. S. Hawley, eds. (Cold Spring Harbor Laboratory Press, 2000), pp. 141–157.

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