Abstract

Tiling light sheet selective plane illumination microscopy (TLS-SPIM) improves the 3D imaging ability of SPIM by using real-time optimized tiling light sheets. However, the imaging speed decreases and the raw image size increases due to the tiling process and additional camera exposures. The decreased imaging speed and the increased raw data could cause significant problems when TLS-SPIM is used to image large specimens at high spatial resolutions. Here, we present a novel method to solve the problem. Discontinuous light sheets created by scanning coaxial beam arrays synchronized with the detection camera rolling shutter are used in TLS-SPIM for 3D imaging. It improves the imaging efficiency of TLS-SPIM by reducing the number of tiles required per image plane without influencing the spatial resolution. We investigate the method via numerical simulations and experiments. We demonstrate the imaging ability of the TLS-SPIM using discontinuous light sheets and show the improved imaging efficiency by imaging optically cleared mouse brain.

© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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Lattice light sheet microscopy using tiling lattice light sheets

Liang Gao, Wei-Chun Tang, Yun-Chi Tsai, and Bi-Chang Chen
Opt. Express 27(2) 1497-1506 (2019)

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2019 (1)

2018 (2)

R. M. Power and J. Huisken, “Adaptable, illumination patterning light sheet microscopy,” Sci. Rep. 8(1), 9615 (2018).
[Crossref]

D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
[Crossref]

2017 (2)

S. I. Kubota, K. Takahashi, J. Nishida, Y. Morishita, S. Ehata, K. Tainaka, K. Miyazono, and H. R. Ueda, “Whole-Body Profiling of Cancer Metastasis with Single-Cell Resolution,” Cell Rep. 20(1), 236–250 (2017).
[Crossref]

R. M. Power and J. Huisken, “A guide to light-sheet fluorescence microscopy for multiscale imaging,” Nat. Methods 14(4), 360–373 (2017).
[Crossref]

2016 (2)

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]

Z. Huang, Z. Hao, and L. Zhu, “Axial multifocal binary-phase zone plate for high numerical aperture focusing,” Opt. Eng. 55(12), 123101 (2016).
[Crossref]

2015 (6)

L. Gao, “Extend the field of view of selective plan illumination microscopy by tiling the excitation light sheet,” Opt. Express 23(5), 6102–6111 (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]

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]

A. K. Chmielewski, A. Kyrsting, P. Mahou, M. T. Wayland, L. Muresan, J. F. Evers, and C. F. Kaminski, “Fast imaging of live organisms with sculpted light sheets,” Sci. Rep. 5(1), 9385 (2015).
[Crossref]

R. K. Chhetri, F. Amat, Y. Wan, B. Höckendorf, W. C. Lemon, and P. J. Keller, “Whole-animal functional and developmental imaging with isotropic spatial resolution,” Nat. Methods 12(12), 1171–1178 (2015).
[Crossref]

L. Gao, “Optimization of the excitation light sheet in selective plane illumination microscopy,” Biomed. Opt. Express 6(3), 881–890 (2015).
[Crossref]

2014 (4)

L. Gao, L. Shao, B.-C. Chen, and E. Betzig, “3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy,” Nat. Protoc. 9(5), 1083–1101 (2014).
[Crossref]

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
[Crossref]

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-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]

L. Zhu, M. Sun, M. Zhu, J. Chen, X. Gao, W. Ma, and D. Zhang, “Three-dimensional shape-controllable focal spot array created by focusing vortex beams modulated by multi-value pure-phase grating,” Opt. Express 22(18), 21354–21367 (2014).
[Crossref]

2013 (2)

B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
[Crossref]

Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
[Crossref]

2012 (6)

R. Tomer, K. Khairy, F. Amat, and P. J. Keller, “Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy,” Nat. Methods 9(7), 755–763 (2012).
[Crossref]

U. Krzic, S. Gunther, T. E. Saunders, S. J. Streichan, and L. Hufnagel, “Multiview light-sheet microscope for rapid in toto imaging,” Nat. Methods 9(7), 730–733 (2012).
[Crossref]

L. Gao, L. Shao, C. D. Higgins, J. S. Poulton, M. Peifer, M. W. Davidson, X. Wu, B. Goldstein, and E. Betzig, “Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens,” Cell 151(6), 1370–1385 (2012).
[Crossref]

L. Silvestri, A. Bria, L. Sacconi, G. Iannello, and F. S. Pavone, “Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain,” Opt. Express 20(18), 20582–20598 (2012).
[Crossref]

E. Baumgart and U. Kubitscheck, “Scanned light-sheet microscopy with confocal slit detection,” Opt. Express 20(19), 21805–21814 (2012).
[Crossref]

J. Yu, C. Zhou, W. Jia, W. Cao, S. Wang, J. Ma, and H. Cao, “Three dimensional Dammann array,” Appl. Opt. 51(10), 1619–1630 (2012).
[Crossref]

2011 (1)

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 (1)

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

2007 (1)

Aanstad, P.

B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
[Crossref]

Amat, F.

R. K. Chhetri, F. Amat, Y. Wan, B. Höckendorf, W. C. Lemon, and P. J. Keller, “Whole-animal functional and developmental imaging with isotropic spatial resolution,” Nat. Methods 12(12), 1171–1178 (2015).
[Crossref]

R. Tomer, K. Khairy, F. Amat, and P. J. Keller, “Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy,” Nat. Methods 9(7), 755–763 (2012).
[Crossref]

Bao, Z.

Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
[Crossref]

Baumgart, E.

Bembenek, J. N.

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
[Crossref]

Betzig, E.

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
[Crossref]

L. Gao, L. Shao, B.-C. Chen, and E. Betzig, “3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy,” Nat. Protoc. 9(5), 1083–1101 (2014).
[Crossref]

L. Gao, L. Shao, C. D. Higgins, J. S. Poulton, M. Peifer, M. W. Davidson, X. Wu, B. Goldstein, and E. Betzig, “Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens,” Cell 151(6), 1370–1385 (2012).
[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]

Böhme, R.

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
[Crossref]

Bria, A.

Bugde, A.

D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
[Crossref]

Campos, C. P.

B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
[Crossref]

Cao, H.

Cao, W.

Chen, B.-C.

L. Gao, W.-C. Tang, Y.-C. Tsai, and B.-C. Chen, “Lattice light sheet microscopy using tiling lattice light sheets,” Opt. Express 27(2), 1497–1506 (2019).
[Crossref]

L. Gao, L. Shao, B.-C. Chen, and E. Betzig, “3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy,” Nat. Protoc. 9(5), 1083–1101 (2014).
[Crossref]

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
[Crossref]

Y.-C. Tsai, W.-C. Tang, C. S. L. Low, Y.-T. Liu, J.-S. Wu, P.-Y. Lee, L. Q. Chen, Y.-L. Lin, P. Kanchanawong, L. Gao, and B.-C. Chen, “Rapid high resolution 3D imaging of expanded biological specimens with lattice light sheet microscopy,” Methods, in press.

Chen, J.

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, L. Q.

Y.-C. Tsai, W.-C. Tang, C. S. L. Low, Y.-T. Liu, J.-S. Wu, P.-Y. Lee, L. Q. Chen, Y.-L. Lin, P. Kanchanawong, L. Gao, and B.-C. Chen, “Rapid high resolution 3D imaging of expanded biological specimens with lattice light sheet microscopy,” Methods, in press.

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]

Chhetri, R. K.

R. K. Chhetri, F. Amat, Y. Wan, B. Höckendorf, W. C. Lemon, and P. J. Keller, “Whole-animal functional and developmental imaging with isotropic spatial resolution,” Nat. Methods 12(12), 1171–1178 (2015).
[Crossref]

Chmielewski, A. K.

A. K. Chmielewski, A. Kyrsting, P. Mahou, M. T. Wayland, L. Muresan, J. F. Evers, and C. F. Kaminski, “Fast imaging of live organisms with sculpted light sheets,” Sci. Rep. 5(1), 9385 (2015).
[Crossref]

Christensen, R.

Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
[Crossref]

Cižmár, T.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-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]

Coll-Lladó, C.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-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]

Colón-Ramos, D. A.

Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
[Crossref]

Dalgarno, H. I. C.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-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]

Danuser, G.

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]

Davidson, M. W.

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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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Y.-C. Tsai, W.-C. Tang, C. S. L. Low, Y.-T. Liu, J.-S. Wu, P.-Y. Lee, L. Q. Chen, Y.-L. Lin, P. Kanchanawong, L. Gao, and B.-C. Chen, “Rapid high resolution 3D imaging of expanded biological specimens with lattice light sheet microscopy,” Methods, in press.

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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
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[Crossref]

Ma, J.

Ma, W.

Mahou, P.

A. K. Chmielewski, A. Kyrsting, P. Mahou, M. T. Wayland, L. Muresan, J. F. Evers, and C. F. Kaminski, “Fast imaging of live organisms with sculpted light sheets,” Sci. Rep. 5(1), 9385 (2015).
[Crossref]

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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]

Matus, D. Q.

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).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
[Crossref]

Milkie, D. E.

B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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S. I. Kubota, K. Takahashi, J. Nishida, Y. Morishita, S. Ehata, K. Tainaka, K. Miyazono, and H. R. Ueda, “Whole-Body Profiling of Cancer Metastasis with Single-Cell Resolution,” Cell Rep. 20(1), 236–250 (2017).
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S. I. Kubota, K. Takahashi, J. Nishida, Y. Morishita, S. Ehata, K. Tainaka, K. Miyazono, and H. R. Ueda, “Whole-Body Profiling of Cancer Metastasis with Single-Cell Resolution,” Cell Rep. 20(1), 236–250 (2017).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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A. K. Chmielewski, A. Kyrsting, P. Mahou, M. T. Wayland, L. Muresan, J. F. Evers, and C. F. Kaminski, “Fast imaging of live organisms with sculpted light sheets,” Sci. Rep. 5(1), 9385 (2015).
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T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-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).
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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).
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L. Gao, L. Shao, C. D. Higgins, J. S. Poulton, M. Peifer, M. W. Davidson, X. Wu, B. Goldstein, and E. Betzig, “Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens,” Cell 151(6), 1370–1385 (2012).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
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F. O. Fahrbach, P. Simon, and A. Rohrbach, “Microscopy with self-reconstructing beams,” Nat. Photonics 4(11), 780–785 (2010).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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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).
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B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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L. Gao, L. Shao, C. D. Higgins, J. S. Poulton, M. Peifer, M. W. Davidson, X. Wu, B. Goldstein, and E. Betzig, “Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens,” Cell 151(6), 1370–1385 (2012).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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U. Krzic, S. Gunther, T. E. Saunders, S. J. Streichan, and L. Hufnagel, “Multiview light-sheet microscope for rapid in toto imaging,” Nat. Methods 9(7), 730–733 (2012).
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Sun, Y.

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).
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S. I. Kubota, K. Takahashi, J. Nishida, Y. Morishita, S. Ehata, K. Tainaka, K. Miyazono, and H. R. Ueda, “Whole-Body Profiling of Cancer Metastasis with Single-Cell Resolution,” Cell Rep. 20(1), 236–250 (2017).
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B. Schmid, G. Shah, N. Scherf, M. Weber, K. Thierbach, C. P. Campos, I. Roeder, P. Aanstad, and J. Huisken, “High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics,” Nat. Commun. 4(1), 2207 (2013).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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S. I. Kubota, K. Takahashi, J. Nishida, Y. Morishita, S. Ehata, K. Tainaka, K. Miyazono, and H. R. Ueda, “Whole-Body Profiling of Cancer Metastasis with Single-Cell Resolution,” Cell Rep. 20(1), 236–250 (2017).
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B.-C. Chen, W. R. Legant, K. Wang, L. Shao, D. E. Milkie, M. W. Davidson, C. Janetopoulos, X. S. Wu, J. A. Hammer, Z. Liu, B. P. English, Y. Mimori-Kiyosue, D. P. Romero, A. T. Ritter, J. Lippincott-Schwartz, L. Fritz-Laylin, R. D. Mullins, D. M. Mitchell, J. N. Bembenek, A. C. Reymann, R. Böhme, S. W. Grill, J. T. Wang, G. Seydoux, U. S. Tulu, D. P. Kiehart, and E. Betzig, “Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution,” Science 346(6208), 1257998 (2014).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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A. K. Chmielewski, A. Kyrsting, P. Mahou, M. T. Wayland, L. Muresan, J. F. Evers, and C. F. Kaminski, “Fast imaging of live organisms with sculpted light sheets,” Sci. Rep. 5(1), 9385 (2015).
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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).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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L. Gao, L. Shao, C. D. Higgins, J. S. Poulton, M. Peifer, M. W. Davidson, X. Wu, B. Goldstein, and E. Betzig, “Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens,” Cell 151(6), 1370–1385 (2012).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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Y. Wu, P. Wawrzusin, J. Senseney, R. S. Fischer, R. Christensen, A. Santella, A. G. York, P. W. Winter, C. M. Waterman, Z. Bao, D. A. Colón-Ramos, M. McAuliffe, and H. Shroff, “Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy,” Nat. Biotechnol. 31(11), 1032–1038 (2013).
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Zhang, D.

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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).
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D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
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D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, J. Q. Feng, L. Gao, W.-P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
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Zhou, Z.

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).
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Appl. Opt. (1)

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Figures (7)

Fig. 1.
Fig. 1. The concept and operation modes of using discontinuous light sheets in TLS-SPIM for 3D imaging. (a) A discontinuous light sheet with multiple waists images a larger effective area at each tile than that of using a continuous light sheet with a single waist. It improves the imaging speed and decreases the raw data size at the same time. (b) The imaging plane is imaged by tiling the same discontinuous light sheet at multiple positions. (c) The imaging plane is imaged by using multiple different discontinuous light sheets compensating each other.
Fig. 2.
Fig. 2. The pupil segmentation method used to create coaxial beam arrays in TLS-SPIM. (a) One group of pupil segments and a binary phase map used to generate a pair of beams within a four-beam coaxial beam array. (b) The other group of pupil segments and the binary phase map used to generate the second pair of coaxial beams within a four-beam coaxial beam array. (c) The combined phase map used to generate the desired four-beam coaxial beam array. (d) The binary phase map used to tile the obtained four-beam coaxial beam array at a different position. Excitation numerical aperture (NA): NAod=0.08, NAid=0.03. (e, f) The intensity profile of the coaxial beam array in Fig. 2(c) at the indicated positions.
Fig. 3.
Fig. 3. The intensity profile comparison of discontinuous light sheets created by scanning coaxial beam arrays and Bessel light sheets created by scanning Bessel beams. (a) YZ max intensity projection of a discontinuous light sheet created by scanning the four-beam coaxial beam array in Fig. 2(c), and its intensity profile at the indicated positions. (b) YZ max intensity projection of a Bessel light sheet that has a comparable thickness and effective length with the discontinuous light sheet in Fig. 3(a), and its intensity profile at the indicated position. Excitation numerical NA: NAod=0.08, NAid=0.07. (c) YZ max intensity projection of the equivalent light sheet when the scanning beam array is synchronized with a 7.5 µm wide virtual confocal slit, and its intensity profiles at the indicated position using confocal slits of different widths. (d) YZ max intensity projection of the equivalent light sheet when the scanning Bessel beam is synchronized with a 7.5 µm wide confocal slit, and its intensity profile at the indicated position using confocal slits of different widths.
Fig. 4.
Fig. 4. The TLS microscope designed for cleared tissue imaging. (a) The schematic diagram of the TLS microscope. (b) The optomechanical design of the TLS microscope.
Fig. 5.
Fig. 5. The imaging performance comparison of TLS-SPIM using continuous and discontinuous light sheets. (a, b) XY and YZ max intensity projection of a sample volume imaged using a continuous light sheet created by scanning the excitation beam. Excitation NA: NAod=0.045, NAid=0.015. (c, d) XY and YZ max intensity projection of the same sample volume imaged using the same light sheet with the scanning beam synchronized with a 6 µm wide virtual confocal slit. (e-j) XY and YZ max intensity projections of the same sample volume imaged using discontinuous light sheets created by scanning a two-beam, three-beam and four-beam coaxial beam array synchronized with a 6 µm virtual confocal slit. Excitation NA: NAod=0.08, NAid=0.03 for all discontinuous light sheets. The inserts show zoomed in views of the indicated cells in each panel. Scale bars, 200 µm in Figs. 5(a) and 5(b), 10 µm in all inserts.
Fig. 6.
Fig. 6. The imaging performance comparison of TLS-SPIM using discontinuous light sheets and Bessel light sheets. (a, b) XY and YZ max intensity projections of a sample volume imaged using a continuous light sheet created by scanning the excitation beam synchronized with a 6 µm virtual confocal slit. Excitation NA: NAod=0.08, NAid=0.03. (c, d) XY and YZ max intensity projections of the same sample volume imaged using a twice long Bessel light sheet created by scanning the excitation beam synchronized a 6 µm virtual confocal slit. Excitation NA: NAod=0.08, NAid=0.06. (e, f) XY and YZ max intensity projections of the same sample volume imaged using a three times long Bessel light sheet created by scanning the excitation beam symphonized with a 6 µm virtual confocal slit. Excitation NA: NAod=0.08, NAid=0.067. (g, h) XY and YZ max intensity projections of the same sample volume imaged using a discontinuous light sheet created by scanning a three-beam coaxial beam array synchronized with a 6 µm virtual confocal slit. Excitation NA: NAod=0.08, NAid=0.03. (i-n) XY max intensity projections of the selected image volumes indicated in Figs. 6(a), 6(c), 6(e), 6(g) . (o-t) YZ max intensity projections of the selected image volumes indicated in Figs. 6(b), 6(d), 6(f), 6(h). Scale bars, 200 µm in Figs. 6(a) and 6(b), 20 µm in Figs. 6(i) and 6(o).
Fig. 7.
Fig. 7. Different operation modes using discontinuous light sheets in TLS-SPIM. (a-f) XY and YZ max intensity projections of a sample volume imaged using a three-waists discontinuous light sheet tiled at three positions. Excitation NA: NAod=0.08, NAid=0.03. (g-j) XY and YZ max intensity projections of the same sample volume imaged using a four-waists discontinuous light sheet and a three-waists discontinuous light sheet compensating each other. Excitation NA: NAod=0.08, NAid=0.03 for the four-waists discontinuous light sheet and NAod=0.06, NAid=0.02 for the three-waists discontinuous light sheet. (k, l) XY and YZ max intensity projections of the reconstructed result, which could be obtained using the results shown in Figs. 7(a)–7(f) or Figs. 7(g)–7(j). Scale bars, 200 µm.