Abstract

We present a theoretical investigation of an optical microscope design that achieves wide-field, multiphoton fluorescence microscopy with finer axial resolution than confocal microscopy. Our technique creates a thin plane of excitation light at the sample using height-staggered microlens arrays (HSMAs), wherein the height staggering of microlenses generate temporal focusing to suppress out-of-focus excitation, and the dense spacing of microlenses enables the implementation of structured illumination technique to eliminate residual out-of-focus signal. We use physical optics-based numerical simulations to demonstrate that our proposed technique can achieve diffraction-limited three-dimensional imaging through a simple optical design.

© 2013 OSA

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  1. J. B. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).
  2. W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
    [CrossRef] [PubMed]
  3. J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
    [CrossRef] [PubMed]
  4. D. Oron, E. Tal, and Y. Silberberg, “Scanningless depth-resolved microscopy,” Opt. Express13, 1468–1476 (2005).
    [CrossRef] [PubMed]
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  6. M. Neil, R. Juskaitis, and T. Wilson, “Method of obtaining optical sectioning by using structured light in a conventional microscope,” Opt. Lett.22, 1905–1907 (1997).
    [CrossRef]
  7. B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
    [CrossRef] [PubMed]
  8. I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).
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    [CrossRef]
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    [CrossRef] [PubMed]
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    [CrossRef]
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  17. D. Lim, K. K. Chu, and J. Mertz, “Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy,” Opt. Lett.33, 1819–21 (2008).
    [CrossRef] [PubMed]
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2011 (2)

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination hilo microscopy,” J. Biomed. Opt.16, 016014 (2011).
[CrossRef] [PubMed]

2010 (1)

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

2008 (1)

2006 (1)

2005 (2)

2004 (2)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

2001 (1)

2000 (1)

1998 (1)

1997 (1)

1990 (1)

W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
[CrossRef] [PubMed]

Akira, I.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Andresen, V.

Bao, Z.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Barry, N.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Benedetti, P. A.

Bewersdorf, J.

Blake, G. A.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Born, M.

M. Born and E. Wolf, Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light, 6th ed. (Cambridge University Press, 1997).

Brenner, K.-H.

J. Jahns and K.-H. Brenner, Microoptics: from Technology to Applications, vol. v. 97 (Springer, 2004).

Buehler, C.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Chen, Y.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Chu, K. K.

D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination hilo microscopy,” J. Biomed. Opt.16, 016014 (2011).
[CrossRef] [PubMed]

D. Lim, K. K. Chu, and J. Mertz, “Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy,” Opt. Lett.33, 1819–21 (2008).
[CrossRef] [PubMed]

Del Bene, F.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Denk, W.

W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
[CrossRef] [PubMed]

Egner, A.

Ford, T. N.

D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination hilo microscopy,” J. Biomed. Opt.16, 016014 (2011).
[CrossRef] [PubMed]

Gratton, E.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Guo, C.-L.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Hecht, E.

E. Hecht, Optics, 4th ed. (Addison-Wesley, 2002).

Heintzmann, R.

Hell, S.

Holland, D. B.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Huisken, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Isao, U.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Jahns, J.

J. Jahns and K.-H. Brenner, Microoptics: from Technology to Applications, vol. v. 97 (Springer, 2004).

Juskaitis, R.

Katsumi, I.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Keller, P. J.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Kenta, M.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Khairy, K.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Kuo, C.-H.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Lim, D.

D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination hilo microscopy,” J. Biomed. Opt.16, 016014 (2011).
[CrossRef] [PubMed]

D. Lim, K. K. Chu, and J. Mertz, “Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy,” Opt. Lett.33, 1819–21 (2008).
[CrossRef] [PubMed]

Mantulin, W.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Masters, B.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Mertz, J.

Michio, A.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Neil, M.

Oron, D.

Ouyang, M.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Pawley, J. B.

J. B. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).

Pick, R.

Santella, A.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Schmidt, A. D.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Silberberg, Y.

So, P.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Stelzer, E.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Stelzer, E. H. K.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Strickler, J.

W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
[CrossRef] [PubMed]

Sutin, J.

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

Swoger, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Takeo, T.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Tal, E.

Ventalon, C.

Webb, W.

W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
[CrossRef] [PubMed]

Wilson, T.

Wittbrodt, J.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Wolf, E.

M. Born and E. Wolf, Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light, 6th ed. (Cambridge University Press, 1997).

Yasuhito, K.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Yu, J.-Y.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Yumiko, S.

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

Zadoyan, R.

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

Appl. Opt. (1)

Bioimaging (1)

I. Akira, T. Takeo, I. Katsumi, S. Yumiko, K. Yasuhito, M. Kenta, A. Michio, and U. Isao, “High-speed confocal fluorescence microscopy using a nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimaging4, 57–62 (1996-06).

J. Biomed. Opt. (3)

B. Masters, P. So, C. Buehler, N. Barry, J. Sutin, W. Mantulin, and E. Gratton, “Mitigating thermal mechanical damage potential during two-photon dermal imaging,” J. Biomed. Opt.9, 1265–1270 (2004).
[CrossRef] [PubMed]

J.-Y. Yu, C.-H. Kuo, D. B. Holland, Y. Chen, M. Ouyang, G. A. Blake, R. Zadoyan, and C.-L. Guo, “Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy,” J. Biomed. Opt.16, 116009 (2011).
[CrossRef] [PubMed]

D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination hilo microscopy,” J. Biomed. Opt.16, 016014 (2011).
[CrossRef] [PubMed]

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

Nat. Methods (1)

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. K. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7, 637–U55 (2010).
[CrossRef] [PubMed]

Opt. Express (1)

Opt. Lett. (5)

Science (2)

W. Denk, J. Strickler, and W. Webb, “2-photon laser scanning fluorescence microscopy,” Science248, 73–76 (1990).
[CrossRef] [PubMed]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science305, 1007–1009 (2004).
[CrossRef] [PubMed]

Other (5)

T. Wilson, Confocal Microscopy (Academic Press, 1990).

J. B. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).

J. Jahns and K.-H. Brenner, Microoptics: from Technology to Applications, vol. v. 97 (Springer, 2004).

M. Born and E. Wolf, Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light, 6th ed. (Cambridge University Press, 1997).

E. Hecht, Optics, 4th ed. (Addison-Wesley, 2002).

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