Abstract

We demonstrate the first planar Airy light-sheet microscope. Fluorescence light-sheet microscopy has become the method of choice to study large biological samples with cellular or sub-cellular resolution. The propagation-invariant Airy beam enables a ten-fold increase in field-of-view with single-photon excitation; however, the characteristic asymmetry of the light-sheet limits its potential for multi-photon excitation. Here we show how a planar light-sheet can be formed from the curved propagation-invariant Airy beam. The resulting symmetric light sheet excites two-photon fluorescence uniformly across an extended field-of-view without the need for deconvolution. We demonstrate the method for rapid two-photon imaging of large volumes of neuronal tissue.

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  1. E. M. Hillman, V. Voleti, W. Li, and H. Yu, “Light-sheet microscopy in neuroscience,” Annu. Rev. Neurosci. 42(1), 295–313 (2019). PMID: 31283896.
    [Crossref]
  2. J. Huisken, J. Swoger, F. D. Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004).
    [Crossref]
  3. M. B. Ahrens, M. B. Orger, D. N. Robson, J. M. Li, and P. J. Keller, “Whole-brain functional imaging at cellular resolution using light-sheet microscopy,” Nat. Methods 10(5), 413–420 (2013).
    [Crossref]
  4. Q. Fu, B. L. Martin, D. Q. Matus, and L. Gao, “Imaging multicellular specimens with real-time optimized tiling light-sheet selective plane illumination microscopy,” Nat. Commun. 7(1), 11088 (2016).
    [Crossref]
  5. K. M. Dean, P. Roudot, E. S. Welf, G. Danuser, and R. Fiolka, “Deconvolution-free subcellular imaging with axially swept light sheet microscopy,” Biophys. J. 108(12), 2807–2815 (2015).
    [Crossref]
  6. T. Chakraborty, M. K. Driscoll, E. Jeffery, M. M. Murphy, P. Roudot, B.-J. Chang, S. Vora, W. M. Wong, C. D. Nielson, H. Zhang, V. Zhemkov, C. Hiremath, E. D. De La Cruz, Y. Yi, I. Bezprozvanny, H. Zhao, R. Tomer, R. Heintzmann, J. P. Meeks, D. K. Marciano, S. J. Morrison, G. Danuser, K. M. Dean, and R. Fiolka, “Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution,” Nat. Methods 16(11), 1109–1113 (2019).
    [Crossref]
  7. F. O. Fahrbach, P. Simon, and A. Rohrbach, “Microscopy with self-reconstructing beams,” Nat. Photonics 4(11), 780–785 (2010).
    [Crossref]
  8. T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
    [Crossref]
  9. T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. C. Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
    [Crossref]
  10. Z. Yang, M. Prokopas, J. Nylk, C. Coll-Lladó, F. J. Gunn-Moore, D. E. K. Ferrier, T. Vettenburg, and K. Dholakia, “A compact Airy beam light sheet microscope with a tilted cylindrical lens,” Biomed. Opt. Express 5(10), 3434–3442 (2014).
    [Crossref]
  11. O. E. Olarte, J. Andilla, D. Artigas, and P. Loza-Alvarez, “Decoupled illumination detection in light sheet microscopy for fast volumetric imaging,” Optica 2(8), 702–705 (2015).
    [Crossref]
  12. T. V. Truong, W. Supatto, D. S. Koos, J. M. Choi, and S. E. Fraser, “Deep and fast live imaging with two-photon scanned light-sheet microscopy,” Nat. Methods 8(9), 757–760 (2011).
    [Crossref]
  13. O. E. Olarte, J. Licea-Rodriguez, J. A. Palero, E. J. Gualda, D. Artigas, J. Mayer, J. Swoger, J. Sharpe, I. Rocha-Mendoza, R. Rangel-Rojo, and P. Loza-Alvarez, “Image formation by linear and nonlinear digital scanned light-sheet fluorescence microscopy with Gaussian and Bessel beam profiles,” Biomed. Opt. Express 3(7), 1492–1505 (2012).
    [Crossref]
  14. Z. Lavagnino, F. C. Zanacchi, E. Ronzitti, and A. Diaspro, “Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application,” Opt. Express 21(5), 5998–6008 (2013).
    [Crossref]
  15. W. Zong, J. Zhao, X. Chen, Y. Lin, H. Ren, Y. Zhang, M. Fan, Z. Zhou, H. Cheng, Y. Sun, and L. Chen, “Large-field high-resolution two-photon digital scanned light-sheet microscopy,” Cell Res. 25(2), 254–257 (2015).
    [Crossref]
  16. S. Wolf, W. Supatto, G. Debregeas, P. Mahou, S. G. Kruglik, J.-M. Sintes, E. Beaurepaire, and R. Candelier, “Whole-brain functional imaging with two-photon light-sheet microscopy,” Nat. Methods 12(5), 379–380 (2015).
    [Crossref]
  17. P. Piksarv, D. Marti, T. Le, A. Unterhuber, L. H. Forbes, M. R. Andrews, A. Stingl, W. Drexler, P. E. Andersen, and K. Dholakia, “Integrated single-and two-photon light sheet microscopy using accelerating beams,” Sci. Rep. 7(1), 1435 (2017).
    [Crossref]
  18. X.-J. Tan, C. Kong, Y.-X. Ren, C. S. W. Lai, K. K. Tsia, and K. K. Y. Wong, “Volumetric two-photon microscopy with a non-diffracting Airy beam,” Opt. Lett. 44(2), 391–394 (2019).
    [Crossref]
  19. G. A. Siviloglou and D. N. Christodoulides, “Accelerating finite energy Airy beams,” Opt. Lett. 32(8), 979–981 (2007).
    [Crossref]
  20. G. A. Siviloglou, J. Broky, A. Dogariu, and D. N. Christodoulides, “Observation of accelerating Airy beams,” Phys. Rev. Lett. 99(21), 213901 (2007).
    [Crossref]
  21. A. D. Jeffrey, J. M. Mark, M. A. Bandres, and D. M. Cottrell, “Observation of accelerating parabolic beams,” Opt. Express 16(17), 12866–12871 (2008).
    [Crossref]
  22. P. J. Keller and E. H. K. Stelzer, “Quantitative in vivo imaging of entire embryos with digital scanned laser light sheet fluorescence microscopy,” Curr. Opin. Neurobiol. 18(6), 624–632 (2008).
    [Crossref]
  23. M. V. Berry and N. L. Balazs, “Nonspreading wave packets,” Am. J. Phys. 47(3), 264–267 (1979).
    [Crossref]
  24. G. Porat, I. Dolev, O. Barlev, and A. Arie, “Airy beam laser,” Opt. Lett. 36(20), 4119–4121 (2011).
    [Crossref]
  25. J. Wang, J. Bu, M. Wang, Y. Yang, and X. Yuan, “Generation of high quality airy beams with blazed micro-optical cubic phase plates,” Appl. Opt. 50(36), 6627–6631 (2011).
    [Crossref]
  26. Y. Wu, A. Ghitani, R. Christensen, A. Santella, Z. Du, G. Rondeau, Z. Bao, D. Colón-Ramos, and H. Shroff, “Inverted selective plane illumination microscopy (ispim) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans,” Proc. Natl. Acad. Sci. U. S. A. 108(43), 17708–17713 (2011).
    [Crossref]
  27. D. G. Papazoglou, S. Suntsov, D. Abdollahpour, and S. Tzortzakis, “Tunable intense Airy beams and tailored femtosecond laser filaments,” Phys. Rev. A 81(6), 061807 (2010).
    [Crossref]
  28. A. Escobet-Montalbán, F. M. Gasparoli, J. Nylk, P. Liu, Z. Yang, and K. Dholakia, “Three-photon light-sheet fluorescence microscopy,” Opt. Lett. 43(21), 5484–5487 (2018).
    [Crossref]
  29. J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
    [Crossref]
  30. R. Tomer, L. Ye, B. Hsueh, and K. Deisseroth, “Advanced clarity for rapid and high-resolution imaging of intact tissues,” Nat. Protoc. 9(7), 1682–1697 (2014).
    [Crossref]
  31. J. H. Yi, D. H. Kim, T. M. Piers, S. C. Kim, D. J. Whitcomb, P. Regan, and K. Cho, “Postsynaptic p47phox regulates long-term depression in the hippocampus,” Cell Discovery 4(1), 44 (2018).
    [Crossref]
  32. J. H. Yi, C. Brown, G. Whitehead, T. Piers, Y. S. Lee, C. Martinez Perez, P. Regan, D. J. Whitcomb, and K. Cho, “Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus,” Pharmacol. Res. 121, 42–51 (2017).
    [Crossref]

2019 (3)

E. M. Hillman, V. Voleti, W. Li, and H. Yu, “Light-sheet microscopy in neuroscience,” Annu. Rev. Neurosci. 42(1), 295–313 (2019). PMID: 31283896.
[Crossref]

T. Chakraborty, M. K. Driscoll, E. Jeffery, M. M. Murphy, P. Roudot, B.-J. Chang, S. Vora, W. M. Wong, C. D. Nielson, H. Zhang, V. Zhemkov, C. Hiremath, E. D. De La Cruz, Y. Yi, I. Bezprozvanny, H. Zhao, R. Tomer, R. Heintzmann, J. P. Meeks, D. K. Marciano, S. J. Morrison, G. Danuser, K. M. Dean, and R. Fiolka, “Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution,” Nat. Methods 16(11), 1109–1113 (2019).
[Crossref]

X.-J. Tan, C. Kong, Y.-X. Ren, C. S. W. Lai, K. K. Tsia, and K. K. Y. Wong, “Volumetric two-photon microscopy with a non-diffracting Airy beam,” Opt. Lett. 44(2), 391–394 (2019).
[Crossref]

2018 (2)

A. Escobet-Montalbán, F. M. Gasparoli, J. Nylk, P. Liu, Z. Yang, and K. Dholakia, “Three-photon light-sheet fluorescence microscopy,” Opt. Lett. 43(21), 5484–5487 (2018).
[Crossref]

J. H. Yi, D. H. Kim, T. M. Piers, S. C. Kim, D. J. Whitcomb, P. Regan, and K. Cho, “Postsynaptic p47phox regulates long-term depression in the hippocampus,” Cell Discovery 4(1), 44 (2018).
[Crossref]

2017 (2)

J. H. Yi, C. Brown, G. Whitehead, T. Piers, Y. S. Lee, C. Martinez Perez, P. Regan, D. J. Whitcomb, and K. Cho, “Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus,” Pharmacol. Res. 121, 42–51 (2017).
[Crossref]

P. Piksarv, D. Marti, T. Le, A. Unterhuber, L. H. Forbes, M. R. Andrews, A. Stingl, W. Drexler, P. E. Andersen, and K. Dholakia, “Integrated single-and two-photon light sheet microscopy using accelerating beams,” Sci. Rep. 7(1), 1435 (2017).
[Crossref]

2016 (1)

Q. Fu, B. L. Martin, D. Q. Matus, and L. Gao, “Imaging multicellular specimens with real-time optimized tiling light-sheet selective plane illumination microscopy,” Nat. Commun. 7(1), 11088 (2016).
[Crossref]

2015 (4)

K. M. Dean, P. Roudot, E. S. Welf, G. Danuser, and R. Fiolka, “Deconvolution-free subcellular imaging with axially swept light sheet microscopy,” Biophys. J. 108(12), 2807–2815 (2015).
[Crossref]

O. E. Olarte, J. Andilla, D. Artigas, and P. Loza-Alvarez, “Decoupled illumination detection in light sheet microscopy for fast volumetric imaging,” Optica 2(8), 702–705 (2015).
[Crossref]

W. Zong, J. Zhao, X. Chen, Y. Lin, H. Ren, Y. Zhang, M. Fan, Z. Zhou, H. Cheng, Y. Sun, and L. Chen, “Large-field high-resolution two-photon digital scanned light-sheet microscopy,” Cell Res. 25(2), 254–257 (2015).
[Crossref]

S. Wolf, W. Supatto, G. Debregeas, P. Mahou, S. G. Kruglik, J.-M. Sintes, E. Beaurepaire, and R. Candelier, “Whole-brain functional imaging with two-photon light-sheet microscopy,” Nat. Methods 12(5), 379–380 (2015).
[Crossref]

2014 (3)

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. C. Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref]

Z. Yang, M. Prokopas, J. Nylk, C. Coll-Lladó, F. J. Gunn-Moore, D. E. K. Ferrier, T. Vettenburg, and K. Dholakia, “A compact Airy beam light sheet microscope with a tilted cylindrical lens,” Biomed. Opt. Express 5(10), 3434–3442 (2014).
[Crossref]

R. Tomer, L. Ye, B. Hsueh, and K. Deisseroth, “Advanced clarity for rapid and high-resolution imaging of intact tissues,” Nat. Protoc. 9(7), 1682–1697 (2014).
[Crossref]

2013 (2)

Z. Lavagnino, F. C. Zanacchi, E. Ronzitti, and A. Diaspro, “Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application,” Opt. Express 21(5), 5998–6008 (2013).
[Crossref]

M. B. Ahrens, M. B. Orger, D. N. Robson, J. M. Li, and P. J. Keller, “Whole-brain functional imaging at cellular resolution using light-sheet microscopy,” Nat. Methods 10(5), 413–420 (2013).
[Crossref]

2012 (2)

O. E. Olarte, J. Licea-Rodriguez, J. A. Palero, E. J. Gualda, D. Artigas, J. Mayer, J. Swoger, J. Sharpe, I. Rocha-Mendoza, R. Rangel-Rojo, and P. Loza-Alvarez, “Image formation by linear and nonlinear digital scanned light-sheet fluorescence microscopy with Gaussian and Bessel beam profiles,” Biomed. Opt. Express 3(7), 1492–1505 (2012).
[Crossref]

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[Crossref]

2011 (5)

G. Porat, I. Dolev, O. Barlev, and A. Arie, “Airy beam laser,” Opt. Lett. 36(20), 4119–4121 (2011).
[Crossref]

J. Wang, J. Bu, M. Wang, Y. Yang, and X. Yuan, “Generation of high quality airy beams with blazed micro-optical cubic phase plates,” Appl. Opt. 50(36), 6627–6631 (2011).
[Crossref]

Y. Wu, A. Ghitani, R. Christensen, A. Santella, Z. Du, G. Rondeau, Z. Bao, D. Colón-Ramos, and H. Shroff, “Inverted selective plane illumination microscopy (ispim) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans,” Proc. Natl. Acad. Sci. U. S. A. 108(43), 17708–17713 (2011).
[Crossref]

T. V. Truong, W. Supatto, D. S. Koos, J. M. Choi, and S. E. Fraser, “Deep and fast live imaging with two-photon scanned light-sheet microscopy,” Nat. Methods 8(9), 757–760 (2011).
[Crossref]

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref]

2010 (2)

F. O. Fahrbach, P. Simon, and A. Rohrbach, “Microscopy with self-reconstructing beams,” Nat. Photonics 4(11), 780–785 (2010).
[Crossref]

D. G. Papazoglou, S. Suntsov, D. Abdollahpour, and S. Tzortzakis, “Tunable intense Airy beams and tailored femtosecond laser filaments,” Phys. Rev. A 81(6), 061807 (2010).
[Crossref]

2008 (2)

A. D. Jeffrey, J. M. Mark, M. A. Bandres, and D. M. Cottrell, “Observation of accelerating parabolic beams,” Opt. Express 16(17), 12866–12871 (2008).
[Crossref]

P. J. Keller and E. H. K. Stelzer, “Quantitative in vivo imaging of entire embryos with digital scanned laser light sheet fluorescence microscopy,” Curr. Opin. Neurobiol. 18(6), 624–632 (2008).
[Crossref]

2007 (2)

G. A. Siviloglou and D. N. Christodoulides, “Accelerating finite energy Airy beams,” Opt. Lett. 32(8), 979–981 (2007).
[Crossref]

G. A. Siviloglou, J. Broky, A. Dogariu, and D. N. Christodoulides, “Observation of accelerating Airy beams,” Phys. Rev. Lett. 99(21), 213901 (2007).
[Crossref]

2004 (1)

J. Huisken, J. Swoger, F. D. Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004).
[Crossref]

1979 (1)

M. V. Berry and N. L. Balazs, “Nonspreading wave packets,” Am. J. Phys. 47(3), 264–267 (1979).
[Crossref]

Abdollahpour, D.

D. G. Papazoglou, S. Suntsov, D. Abdollahpour, and S. Tzortzakis, “Tunable intense Airy beams and tailored femtosecond laser filaments,” Phys. Rev. A 81(6), 061807 (2010).
[Crossref]

Ahrens, M. B.

M. B. Ahrens, M. B. Orger, D. N. Robson, J. M. Li, and P. J. Keller, “Whole-brain functional imaging at cellular resolution using light-sheet microscopy,” Nat. Methods 10(5), 413–420 (2013).
[Crossref]

Andersen, P. E.

P. Piksarv, D. Marti, T. Le, A. Unterhuber, L. H. Forbes, M. R. Andrews, A. Stingl, W. Drexler, P. E. Andersen, and K. Dholakia, “Integrated single-and two-photon light sheet microscopy using accelerating beams,” Sci. Rep. 7(1), 1435 (2017).
[Crossref]

Andilla, J.

Andrews, M. R.

P. Piksarv, D. Marti, T. Le, A. Unterhuber, L. H. Forbes, M. R. Andrews, A. Stingl, W. Drexler, P. E. Andersen, and K. Dholakia, “Integrated single-and two-photon light sheet microscopy using accelerating beams,” Sci. Rep. 7(1), 1435 (2017).
[Crossref]

Arganda-Carreras, I.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[Crossref]

Arie, A.

Artigas, D.

Balazs, N. L.

M. V. Berry and N. L. Balazs, “Nonspreading wave packets,” Am. J. Phys. 47(3), 264–267 (1979).
[Crossref]

Bandres, M. A.

Bao, Z.

Y. Wu, A. Ghitani, R. Christensen, A. Santella, Z. Du, G. Rondeau, Z. Bao, D. Colón-Ramos, and H. Shroff, “Inverted selective plane illumination microscopy (ispim) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans,” Proc. Natl. Acad. Sci. U. S. A. 108(43), 17708–17713 (2011).
[Crossref]

Barlev, O.

Beaurepaire, E.

S. Wolf, W. Supatto, G. Debregeas, P. Mahou, S. G. Kruglik, J.-M. Sintes, E. Beaurepaire, and R. Candelier, “Whole-brain functional imaging with two-photon light-sheet microscopy,” Nat. Methods 12(5), 379–380 (2015).
[Crossref]

Bene, F. D.

J. Huisken, J. Swoger, F. D. Bene, J. Wittbrodt, and E. H. K. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004).
[Crossref]

Berry, M. V.

M. V. Berry and N. L. Balazs, “Nonspreading wave packets,” Am. J. Phys. 47(3), 264–267 (1979).
[Crossref]

Betzig, E.

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref]

Bezprozvanny, I.

T. Chakraborty, M. K. Driscoll, E. Jeffery, M. M. Murphy, P. Roudot, B.-J. Chang, S. Vora, W. M. Wong, C. D. Nielson, H. Zhang, V. Zhemkov, C. Hiremath, E. D. De La Cruz, Y. Yi, I. Bezprozvanny, H. Zhao, R. Tomer, R. Heintzmann, J. P. Meeks, D. K. Marciano, S. J. Morrison, G. Danuser, K. M. Dean, and R. Fiolka, “Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution,” Nat. Methods 16(11), 1109–1113 (2019).
[Crossref]

Broky, J.

G. A. Siviloglou, J. Broky, A. Dogariu, and D. N. Christodoulides, “Observation of accelerating Airy beams,” Phys. Rev. Lett. 99(21), 213901 (2007).
[Crossref]

Brown, C.

J. H. Yi, C. Brown, G. Whitehead, T. Piers, Y. S. Lee, C. Martinez Perez, P. Regan, D. J. Whitcomb, and K. Cho, “Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus,” Pharmacol. Res. 121, 42–51 (2017).
[Crossref]

Bu, J.

Candelier, R.

S. Wolf, W. Supatto, G. Debregeas, P. Mahou, S. G. Kruglik, J.-M. Sintes, E. Beaurepaire, and R. Candelier, “Whole-brain functional imaging with two-photon light-sheet microscopy,” Nat. Methods 12(5), 379–380 (2015).
[Crossref]

Cardona, A.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods 9(7), 676–682 (2012).
[Crossref]

Chakraborty, T.

T. Chakraborty, M. K. Driscoll, E. Jeffery, M. M. Murphy, P. Roudot, B.-J. Chang, S. Vora, W. M. Wong, C. D. Nielson, H. Zhang, V. Zhemkov, C. Hiremath, E. D. De La Cruz, Y. Yi, I. Bezprozvanny, H. Zhao, R. Tomer, R. Heintzmann, J. P. Meeks, D. K. Marciano, S. J. Morrison, G. Danuser, K. M. Dean, and R. Fiolka, “Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution,” Nat. Methods 16(11), 1109–1113 (2019).
[Crossref]

Chang, B.-J.

T. Chakraborty, M. K. Driscoll, E. Jeffery, M. M. Murphy, P. Roudot, B.-J. Chang, S. Vora, W. M. Wong, C. D. Nielson, H. Zhang, V. Zhemkov, C. Hiremath, E. D. De La Cruz, Y. Yi, I. Bezprozvanny, H. Zhao, R. Tomer, R. Heintzmann, J. P. Meeks, D. K. Marciano, S. J. Morrison, G. Danuser, K. M. Dean, and R. Fiolka, “Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution,” Nat. Methods 16(11), 1109–1113 (2019).
[Crossref]

Chen, L.

W. Zong, J. Zhao, X. Chen, Y. Lin, H. Ren, Y. Zhang, M. Fan, Z. Zhou, H. Cheng, Y. Sun, and L. Chen, “Large-field high-resolution two-photon digital scanned light-sheet microscopy,” Cell Res. 25(2), 254–257 (2015).
[Crossref]

Chen, X.

W. Zong, J. Zhao, X. Chen, Y. Lin, H. Ren, Y. Zhang, M. Fan, Z. Zhou, H. Cheng, Y. Sun, and L. Chen, “Large-field high-resolution two-photon digital scanned light-sheet microscopy,” Cell Res. 25(2), 254–257 (2015).
[Crossref]

Cheng, H.

W. Zong, J. Zhao, X. Chen, Y. Lin, H. Ren, Y. Zhang, M. Fan, Z. Zhou, H. Cheng, Y. Sun, and L. Chen, “Large-field high-resolution two-photon digital scanned light-sheet microscopy,” Cell Res. 25(2), 254–257 (2015).
[Crossref]

Cho, K.

J. H. Yi, D. H. Kim, T. M. Piers, S. C. Kim, D. J. Whitcomb, P. Regan, and K. Cho, “Postsynaptic p47phox regulates long-term depression in the hippocampus,” Cell Discovery 4(1), 44 (2018).
[Crossref]

J. H. Yi, C. Brown, G. Whitehead, T. Piers, Y. S. Lee, C. Martinez Perez, P. Regan, D. J. Whitcomb, and K. Cho, “Glucocorticoids activate a synapse weakening pathway culminating in tau phosphorylation in the hippocampus,” Pharmacol. Res. 121, 42–51 (2017).
[Crossref]

Choi, J. M.

T. V. Truong, W. Supatto, D. S. Koos, J. M. Choi, and S. E. Fraser, “Deep and fast live imaging with two-photon scanned light-sheet microscopy,” Nat. Methods 8(9), 757–760 (2011).
[Crossref]

Christensen, R.

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Supplementary Material (1)

NameDescription
» Visualization 1       Three-dimensional visualization with rotating perspective of 0.60x1.15x0.60mm of Thy-1-GFP-expressing mouse brain tissue, imaged using the two-photon planar Airy light-sheet.

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Figures (4)

Fig. 1.
Fig. 1. Depiction of 2PE light-sheet formation by rapidly scanning a truncated Gaussian beam (A-C), an Airy beam with aligned principal axes (D-F), and an Airy beam with its principal axes at $45^{\circ }$ to the light-sheet (G-I). The detection objective (not shown) is focused to the $x$-$y$-plane and two-photon excited fluorescence is collected along the $z$-axis. Transversal ($y$-$z$) cross-sections are shown in (A,D,G) for the beam waist (magenta) and the 2PE light-sheet (false-color legend in panel (B)). Normalized linear intensity and 2PE for each light-sheet at its waist ($x=0$) are shown in panels (C,F,I). The inset disks show the relative back aperture size and phase (hue). The numerical apertures (NA) for the Airy beams is $0.30$, while that for the truncated Gaussian is chosen to be $0.10$ as a larger NA would have resulted in an impractically narrow field-of-view. The extents of the two-photon light-sheets can be seen in panels (B,E,H), which show false-color $x$-$z$-sections. (B) It can be seen that even at the lower NA, the truncated Gaussian only illuminates a fraction of the full field-of-view. (E) The Airy beam light-sheet illuminates the full field-of-view; however, the side-lobes of the Airy beam can be seen to form parallel, curved, surfaces. As a result, towards the extremes of the field-of-view, the main lobe does not coincide with the focal plane ($z=0$). (H) The $45^{\circ }$-rotated Airy beam can be seen to produce a single, planar, excitation surface. (I) By rotating the scanned beam with respect to the light-sheet plane, its side-lobes merge into a single structure (blue), which results in a highly-confined 2PE (red). Most importantly, the excitation coincides with the focal plane throughout the $0.60\;\textrm{mm}$-wide field-of-view.
Fig. 2.
Fig. 2. Comparison of the two-photon Airy beam light-sheet and the two-photon Planar Airy light-sheet for uniformity and resolution. (A,B) Maximum intensity projection of fluorescent microspheres with two-photon fluorescence excited using the conventional Airy light-sheet (A), and the planar Airy light-sheet (B). (C,D) Intensity profiles of four images of isolated fluorescent microspheres near the center of the field-of-view, $x=0\; \mathrm{\mu}\textrm{m}$ (C, A1, B1) and near $x=-200\; \mathrm{\mu}\textrm{m}$ (D, A2, B2); for the conventional Airy light-sheet (red dashed) and the Planar Airy light-sheet (green solid). The locations of the 3D sections are indicated with green rectangles in panels (A) and (B). All false-color images have been normalized to their maximum intensity for clarity. (E) Peak intensity as a function of distance to the field-of-view center $|x|$ for the microspheres illuminated with the conventional Airy light-sheet (red) and the Planar Airy light-sheet (green). To evaluate the illumination uniformity, normal distributions have been fitted to the intensities and found to have a full-width-at-half-maximum field-of-view of $311\pm 17\; \mathrm{\mu}\textrm{m}$ (Airy, dashed red) and $415\pm 11\; \mathrm{\mu}\textrm{m}$ (Planar Airy, green). (F) Full-width at half maximum in $x$ (dashed), $y$ (dotted), and $z$ (solid) of the microsphere images. Median values for every $10\; \mathrm{\mu}\textrm{m}$ interval are plotted for microspheres with peak intensities at least $50\%$ above the background.
Fig. 3.
Fig. 3. Maximum intensity projection of $0.60 \times 0.60 \times 0.60\;\textrm{mm}^{3}$ of Wistar rat hippocampal tissue, imaged using the two-photon planar Airy light-sheet. (A) The $z$-$y$-projection along the $x$-axis of the volume indicated by the yellow dash-dotted box in panel (B). The variation in axial ($z$) resolution can be seen to be minimal. (B) The $x$-$y$-projection along the $z$-axis, showing high lateral resolution throughout the $0.6\times 0.6\times 0.5\;\textrm{mm}^{3}$ field-of-view. (C) Magnified 3D slice of the data cube, indicated by the orange rectangle in panel (B). (D) A further $5\times$ enlargement of the magnified sub-volume indicated by the orange rectangle in panel (C). White arrowheads indicate examples of dendritic spines.
Fig. 4.
Fig. 4. Maximum intensity projection of $x \times y \times z = 0.60 \times 1.15 \times 0.60\;\textrm{mm}^{3}$ of Thy-1-GFP-expressing mouse brain tissue, imaged using the two-photon planar Airy light-sheet. A video showing additional viewpoints is included as Visualization 1.