Abstract

Conventional, degenerate multiphoton microscopy (D-MPM) requires the use of a high-numerical-aperture (NA) objective. Nondegenerate MPM (ND-MPM) imposes the additional demand for precise spatiotemporal overlap of two distinct excitation sources. We demonstrate that the axial focal shift introduced by refractive objective chromatic aberration hinders the spatial requirement of ND-MPM, whereas the use of a reflective objective overcomes this challenge and allows for improved ND excitation efficiency in spite of a lower NA. Moreover, we demonstrate that reflective objective focusing eliminates the axial misregistration of volumetric stacks in traditional D-MPM experiments when multiple excitation wavelengths are used.

© 2019 Optical Society of America

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References

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    [Crossref]
  2. D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
    [Crossref]
  3. M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
    [Crossref]
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    [Crossref]
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2018 (1)

M. M. Kabir, A. M. Choubal, and K. C. Toussaint, J. Microsc. 271, 129 (2018).
[Crossref]

2017 (2)

B. Amirsolaimani, B. Cromey, N. Peyghambarian, and K. Kieu, Opt. Express 25, 23399 (2017).
[Crossref]

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

2016 (1)

2015 (1)

2013 (1)

2012 (1)

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

2011 (2)

2009 (1)

S. Quentmeier, S. Denicke, and K.-H. Gericke, J. Fluoresc. 19, 1037 (2009).
[Crossref]

2007 (1)

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

2006 (1)

2005 (1)

F. Helmchen and W. Denk, Nat. Methods 2, 932 (2005).
[Crossref]

2004 (1)

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

Abashin, M.

M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
[Crossref]

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

Abdeladim, L.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Adams, S. R.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Amirsolaimani, B.

Beaurepaire, E.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Bovik, A. C.

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

Brizion, S.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Chen, S.

Choubal, A. M.

M. M. Kabir, A. M. Choubal, and K. C. Toussaint, J. Microsc. 271, 129 (2018).
[Crossref]

Cromey, B.

Débarre, D.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Denicke, S.

S. Quentmeier, S. Denicke, and K.-H. Gericke, J. Fluoresc. 19, 1037 (2009).
[Crossref]

Denk, W.

F. Helmchen and W. Denk, Nat. Methods 2, 932 (2005).
[Crossref]

Devor, A.

M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
[Crossref]

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Fainman, Y.

M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
[Crossref]

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

Ferri, C. G.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Ferri, C. G. L.

Fu, D.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

Galey, J.-B.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Ge, N.-H.

Gericke, K.-H.

S. Quentmeier, S. Denicke, and K.-H. Gericke, J. Fluoresc. 19, 1037 (2009).
[Crossref]

Gu, C.

Han, Y.

Helmchen, F.

F. Helmchen and W. Denk, Nat. Methods 2, 932 (2005).
[Crossref]

Jiang, B.

Jiang, J.

Kabir, M. M.

M. M. Kabir, A. M. Choubal, and K. C. Toussaint, J. Microsc. 271, 129 (2018).
[Crossref]

Ke, Y.

Kieu, K.

Kobat, D.

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

Korth, O.

Labroille, G.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Lamarre, I.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Lasser, T.

Legouis, R.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Leitgeb, R. A.

Leutenegger, M.

Levene, M. J.

Liu, Y.

Livet, J.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Loulier, K.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Luo, Q.

Mahou, P.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Malkinson, G.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Matho, K. S.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Matthews, T. E.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

Miller, D. R.

D. R. Miller, “Extending the depth limit of multiphoton microscopy for in vivo brain imaging,” Ph.D. dissertation (The University of Texas at Austin, 2016).

Morin, X.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Pena, A.-M.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Peyghambarian, N.

Potma, E. O.

Quentmeier, S.

S. Quentmeier, S. Denicke, and K.-H. Gericke, J. Fluoresc. 19, 1037 (2009).
[Crossref]

Raghunathan, V.

Rao, R.

Rodriquez, E. A.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Sadegh, S.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Saisan, P. A.

M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
[Crossref]

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

Sheikh, H. R.

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

Simoncelli, E. P.

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

Solinas, X.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Stringari, C.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Supatto, W.

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Thunemann, M.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Tian, P.

Toussaint, K. C.

M. M. Kabir, A. M. Choubal, and K. C. Toussaint, J. Microsc. 271, 129 (2018).
[Crossref]

Vinogradov, S. A.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Walker, S.

Wang, H.

Wang, Z.

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

Warren, W. S.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

Xu, C.

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

Yang, M.-H.

M.-H. Yang, M. Abashin, P. A. Saisan, P. Tian, C. G. L. Ferri, A. Devor, and Y. Fainman, Opt. Express 24, 30173 (2016).
[Crossref]

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

Yang, X.

Ye, T.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

Yung, W. H.

Yurtsever, G.

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

Zhang, D.

Zhu, G.

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

Zimmerley, M.

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Zinter, J. P.

IEEE Trans. Image Process. (1)

Z. Wang, A. C. Bovik, H. R. Sheikh, and E. P. Simoncelli, IEEE Trans. Image Process. 13, 600 (2004).
[Crossref]

J. Biomed. Opt. (1)

D. Fu, T. Ye, T. E. Matthews, G. Yurtsever, and W. S. Warren, J. Biomed. Opt. 12, 054004 (2007).
[Crossref]

J. Fluoresc. (1)

S. Quentmeier, S. Denicke, and K.-H. Gericke, J. Fluoresc. 19, 1037 (2009).
[Crossref]

J. Microsc. (1)

M. M. Kabir, A. M. Choubal, and K. C. Toussaint, J. Microsc. 271, 129 (2018).
[Crossref]

Nat. Methods (2)

F. Helmchen and W. Denk, Nat. Methods 2, 932 (2005).
[Crossref]

P. Mahou, M. Zimmerley, K. Loulier, K. S. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre, and E. Beaurepaire, Nat. Methods 9, 815 (2012).
[Crossref]

Opt. Express (6)

Opt. Lett. (1)

Sci. Rep. (1)

C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, and E. Beaurepaire, Sci. Rep. 7, 3792 (2017).
[Crossref]

Other (4)

D. Kobat, G. Zhu, and C. Xu, in Biomedical Optics (Optical Society of America, 2008), paper BMF6.

M.-H. Yang, M. Abashin, P. A. Saisan, A. Devor, and Y. Fainman, in Optics and the Brain (Optical Society of America, 2015), paper BrW3B.6.

D. R. Miller, “Extending the depth limit of multiphoton microscopy for in vivo brain imaging,” Ph.D. dissertation (The University of Texas at Austin, 2016).

S. Sadegh, M.-H. Yang, C. G. Ferri, M. Thunemann, P. A. Saisan, E. A. Rodriquez, S. R. Adams, S. A. Vinogradov, Y. Fainman, and A. Devor, in Optics and the Brain (Optical Society of America, 2018), paper BF3C.2.

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Figures (4)

Fig. 1.
Fig. 1. (a) Fluorescence intensity curves versus axial focal position characterizes axial focal shift for refractive (i, 10 × ; ii, 20 × ) and reflective (iii, 40 × ) objectives. (b) Axial focal shift values versus excitation wavelength plotted against modeled axial point spread function full width at half-maximum (shaded regions).
Fig. 2.
Fig. 2. Structural-similarity index measurements (SSIM) of individual slices from z -stacks recorded at discrete excitation wavelengths with (a) a 10 × refractive objective, (b) a 20 × refractive objective, and (c) a 40 × reflective objective. For each objective, the SSIM scores were recorded relative to a well-focused reference image from the λ ex = 1150 nm stacks.
Fig. 3.
Fig. 3. Transverse max intensity projections of a cellular monolayer recorded with (a) a 10 × refractive objective, (b) a 20 × refractive objective, and (c) a 40 × reflective objective. (d) Mean image intensity versus depth for the 10 × (dashed lines), 20 × (finely dashed lines), and 40 × (solid lines) objectives. The green channel contains Alexa Fluor 488 signal, λ ex = 790 nm , and the red channel denotes Alexa Fluor 594 signal and second-harmonic generation, λ ex = 1300 nm . Scale bar = 75 μm.
Fig. 4.
Fig. 4. Nondegenerate excitation efficiency comparison of the refractive and reflective objectives; scatter points denote raw data, solid lines indicate regression fits, and shaded regions demarcate corresponding 95% confidence intervals.