Abstract

The intrinsic aberrations of high-NA gradient refractive index (GRIN) lenses limit their image quality as well as field of view. Here we used a pupil-segmentation-based adaptive optical approach to correct the inherent aberrations in a two-photon fluorescence endoscope utilizing a 0.8 NA GRIN lens. By correcting the field-dependent aberrations, we recovered diffraction-limited performance across a large imaging field. The consequent improvements in imaging signal and resolution allowed us to detect fine structures that were otherwise invisible inside mouse brain slices.

© 2012 Optical Society of America

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References

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  1. C. Gomez-Reino, M. V. Perez, and C. Bao, Gradient-Index Optics (Springer-Verlag, 2002).
  2. W. J. Tomlinson, Appl. Opt. 19, 1127 (1980).
    [CrossRef]
  3. J. C. Jung and M. J. Schnitzer, Opt. Lett. 28, 902 (2003).
    [CrossRef]
  4. W. M. Lee and S. H. Yun, Opt. Lett. 36, 4608 (2011).
    [CrossRef]
  5. F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
    [CrossRef]
  6. N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
    [CrossRef]
  7. N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
    [CrossRef]
  8. R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
    [CrossRef]
  9. GRINTECH. Datasheet “High NA for 2-Photon Microscopy,” available from http://www.grintech.de/grin-lens-systems-for-medical-applications.html .

2012 (1)

N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
[CrossRef]

2011 (2)

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

W. M. Lee and S. H. Yun, Opt. Lett. 36, 4608 (2011).
[CrossRef]

2010 (1)

N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
[CrossRef]

2009 (1)

R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
[CrossRef]

2003 (1)

1980 (1)

Bao, C.

C. Gomez-Reino, M. V. Perez, and C. Bao, Gradient-Index Optics (Springer-Verlag, 2002).

Barretto, R. P.

R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
[CrossRef]

Betzig, E.

N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
[CrossRef]

N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
[CrossRef]

Bonoli, C.

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

Bortoletto, F.

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

Ciubotaru, C. D.

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

Gomez-Reino, C.

C. Gomez-Reino, M. V. Perez, and C. Bao, Gradient-Index Optics (Springer-Verlag, 2002).

Ji, N.

N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
[CrossRef]

N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
[CrossRef]

Jung, J. C.

Lee, W. M.

Mammano, F.

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

Messerschmidt, B.

R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
[CrossRef]

Milkie, D. E.

N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
[CrossRef]

Panizzolo, P.

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

Perez, M. V.

C. Gomez-Reino, M. V. Perez, and C. Bao, Gradient-Index Optics (Springer-Verlag, 2002).

Sato, T. R.

N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
[CrossRef]

Schnitzer, M. J.

R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
[CrossRef]

J. C. Jung and M. J. Schnitzer, Opt. Lett. 28, 902 (2003).
[CrossRef]

Tomlinson, W. J.

Yun, S. H.

Appl. Opt. (1)

Nat. Meth. (2)

N. Ji, D. E. Milkie, and E. Betzig, Nat. Meth. 7, 141 (2010).
[CrossRef]

R. P. Barretto, B. Messerschmidt, and M. J. Schnitzer, Nat. Meth. 6, 511 (2009).
[CrossRef]

Opt. Lett. (2)

PLoS ONE (1)

F. Bortoletto, C. Bonoli, P. Panizzolo, C. D. Ciubotaru, and F. Mammano, PLoS ONE 6, e22321 (2011).
[CrossRef]

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

N. Ji, T. R. Sato, and E. Betzig, Proc. Nat. Acad. Sci. USA 109, 22 (2012).
[CrossRef]

Other (2)

GRINTECH. Datasheet “High NA for 2-Photon Microscopy,” available from http://www.grintech.de/grin-lens-systems-for-medical-applications.html .

C. Gomez-Reino, M. V. Perez, and C. Bao, Gradient-Index Optics (Springer-Verlag, 2002).

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Figures (3)

Fig. 1.
Fig. 1.

On-axis AO correction of a 0.8 NA endoscope. (a) Maximal intensity projections of 2 μm diameter fluorescent beads on coverglass before and after AO correction at the center of the FOV. (b) Maximal intensity projections of a brain slice of a YFP line 16 mouse without and with the AO correction in (a). (c) Aberration correction pattern in units of wavelength (λ=900nm). (d), (e) Signal profile along the two dashed lines in (a) and (b), respectively. Scale bars: 20 μm.

Fig. 2.
Fig. 2.

Off-axis AO correction of a 0.8 NA endoscope. (a) Focal series images of two fluorescent beads located at (70.7, 70.7) μm field position before and after AO correction; Images without AO have its brightness digitally enhanced by 8.7× so that both focal series have the same maximal signal. Focal step is 0.5 μm. (b) Similar images to those in (a) obtained at (0, 100) μm. Digital enhancement for images without AO is 6.8×. (c) and (d) are the aberration correction patterns for (a) and (b), respectively, in units of wavelength. (e) Integrated signal of focal series images at different focal planes for 9 field positions: (0,0), (0, 100), (0,100), (100, 0), (100,0), (70.7, 70.7), (70.7, 70.7), (70.7, 70.7), (70.7, 70.7) μm. For each field position, the integrated signal measured without AO is normalized to that measured with AO. Scale bars: 10 μm.

Fig. 3.
Fig. 3.

Enlarged FOV of a 0.8 NA GRIN lens endoscope with AO correction. (a) Maximal intensity projections of 2 μm diameter beads in a 200 μm image field without and with AO corrections; Image with AO correction is the maximal intensity projection of images taken with nine AO correction patterns. (b) Signal profiles along the three dashed lines in (a). (c) Maximal intensity projections of a brain slice before and after AO corrections at nine field positions. (d) Maximal intensity projections of another brain slices at (70.7, 70.7) μm before and after AO correction there. (e) Maximal intensity projections of the brain slice in (c) after AO corrections at opposing field positions (0, 100) and (0, 100) μm. Insets describe the image field positions where AO correction was carried out. Scale bar: 20 μm.

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