Abstract

Neural circuits are fundamental for brain functions. However, obtaining long range continuous projections of neurons in the entire brain is still challenging. Here a two-photon fluorescence micro-optical sectioning tomography (2p-fMOST) method is developed for high-throughput, high-resolution visualization of the brain circuits. Two-photon imaging technology is used to obtain high resolution, and acoustical optical deflector (AOD), an inertia-free beam scanner is used to realize fast and prolonged stable imaging. The combination of these techniques with imaging and then sectioning method of a plastic-embedded mouse brain facilitated the acquisition of a three-dimensional data set of a fluorescent mouse brain with a resolution adequate to resolve the spines. In addition, the brain circuit tracing ability is showed by several neurons projecting across different brain regions. Besides brain imaging, 2p-fMOST could be used in many studies that requires sub-micro resolution or micro resolution imaging of a large sample.

© 2013 OSA

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2013 (2)

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
[CrossRef] [PubMed]

T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
[CrossRef] [PubMed]

2012 (6)

K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS ONE7(3), e33916 (2012).
[CrossRef] [PubMed]

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
[CrossRef] [PubMed]

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

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. Express20(18), 20582–20598 (2012).
[CrossRef] [PubMed]

V. Marx, “High-throughput anatomy: Charting the brain’s networks,” Nature490(7419), 293–298 (2012).
[CrossRef] [PubMed]

O. Eschenko, H. C. Evrard, R. M. Neves, M. Beyerlein, Y. Murayama, and N. K. Logothetis, “Tracing of noradrenergic projections using manganese-enhanced MRI,” Neuroimage59(4), 3252–3265 (2012).
[CrossRef] [PubMed]

2011 (6)

J. Mertz, “Optical sectioning microscopy with planar or structured illumination,” Nat. Methods8(10), 811–819 (2011).
[CrossRef] [PubMed]

J. W. Lichtman and W. Denk, “The big and the small: challenges of imaging the brain’s circuits,” Science334(6056), 618–623 (2011).
[CrossRef] [PubMed]

K. L. Briggman, M. Helmstaedter, and W. Denk, “Wiring specificity in the direction-selectivity circuit of the retina,” Nature471(7337), 183–188 (2011).
[CrossRef] [PubMed]

S. Watanabe, A. Punge, G. Hollopeter, K. I. Willig, R. J. Hobson, M. W. Davis, S. W. Hell, and E. M. Jorgensen, “Protein localization in electron micrographs using fluorescence nanoscopy,” Nat. Methods8(1), 80–84 (2011).
[CrossRef] [PubMed]

X. Liu, T. Quan, S. Zeng, and X. Lv, “Identification of the direction of the neural network activation with a cellular resolution by fast two-photon imaging,” J. Biomed. Opt.16(8), 080506 (2011).
[CrossRef] [PubMed]

X. Chen, U. Leischner, N. L. Rochefort, I. Nelken, and A. Konnerth, “Functional mapping of single spines in cortical neurons in vivo,” Nature475(7357), 501–505 (2011).
[CrossRef] [PubMed]

2010 (5)

B. F. Grewe, D. Langer, H. Kasper, B. M. Kampa, and F. Helmchen, “High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision,” Nat. Methods7(5), 399–405 (2010).
[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

L. A. Harsan, D. Paul, S. Schnell, B. W. Kreher, J. Hennig, J. F. Staiger, and D. von Elverfeldt, “In vivo diffusion tensor magnetic resonance imaging and fiber tracking of the mouse brain,” NMR Biomed.23(7), 884–896 (2010).
[CrossRef] [PubMed]

A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
[CrossRef] [PubMed]

K. Minogue, “Neuroscience. China’s brain mappers zoom in on neural connections,” Science330(6005), 747 (2010).
[CrossRef] [PubMed]

2009 (1)

S. Mori, K. Oishi, and A. V. Faria, “White matter atlases based on diffusion tensor imaging,” Curr. Opin. Neurol.22(4), 362–369 (2009).
[CrossRef] [PubMed]

2008 (4)

S. Canals, M. Beyerlein, A. L. Keller, Y. Murayama, and N. K. Logothetis, “Magnetic resonance imaging of cortical connectivity in vivo,” Neuroimage40(2), 458–472 (2008).
[CrossRef] [PubMed]

L. Luo, E. M. Callaway, and K. Svoboda, “Genetic dissection of neural circuits,” Neuron57(5), 634–660 (2008).
[CrossRef] [PubMed]

G. Duemani Reddy, K. Kelleher, R. Fink, and P. Saggau, “Three-dimensional random access multiphoton microscopy for functional imaging of neuronal activity,” Nat. Neurosci.11(6), 713–720 (2008).
[CrossRef] [PubMed]

D. Mayerich, L. Abbott, and B. McCormick, “Knife-edge scanning microscopy for imaging and reconstruction of three-dimensional anatomical structures of the mouse brain,” J. Microsc.231(1), 134–143 (2008).
[CrossRef] [PubMed]

2007 (4)

D. Li, S. Zeng, X. Lv, J. Liu, R. Du, R. Jiang, W. R. Chen, and Q. Luo, “Dispersion characteristics of acousto-optic deflector for scanning Gaussian laser beam of femtosecond pulses,” Opt. Express15(8), 4726–4734 (2007).
[CrossRef] [PubMed]

K. D. Micheva and S. J. Smith, “Array tomography: a new tool for imaging the molecular architecture and ultrastructure of neural circuits,” Neuron55(1), 25–36 (2007).
[CrossRef] [PubMed]

E. L. Bearer, X. Zhang, and R. E. Jacobs, “Live imaging of neuronal connections by magnetic resonance: Robust transport in the hippocampal-septal memory circuit in a mouse model of Down syndrome,” Neuroimage37(1), 230–242 (2007).
[CrossRef] [PubMed]

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

2006 (1)

2004 (1)

W. Denk and H. Horstmann, “Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure,” PLoS Biol.2(11), e329 (2004).
[CrossRef] [PubMed]

2003 (1)

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

2001 (1)

2000 (1)

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Abbott, L.

D. Mayerich, L. Abbott, and B. McCormick, “Knife-edge scanning microscopy for imaging and reconstruction of three-dimensional anatomical structures of the mouse brain,” J. Microsc.231(1), 134–143 (2008).
[CrossRef] [PubMed]

Arganda-Carreras, I.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

Bahlmann, K.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

Bao, Z.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

Bearer, E. L.

E. L. Bearer, X. Zhang, and R. E. Jacobs, “Live imaging of neuronal connections by magnetic resonance: Robust transport in the hippocampal-septal memory circuit in a mouse model of Down syndrome,” Neuroimage37(1), 230–242 (2007).
[CrossRef] [PubMed]

Becker, K.

K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS ONE7(3), e33916 (2012).
[CrossRef] [PubMed]

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Bernstein, M.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Beyerlein, M.

O. Eschenko, H. C. Evrard, R. M. Neves, M. Beyerlein, Y. Murayama, and N. K. Logothetis, “Tracing of noradrenergic projections using manganese-enhanced MRI,” Neuroimage59(4), 3252–3265 (2012).
[CrossRef] [PubMed]

S. Canals, M. Beyerlein, A. L. Keller, Y. Murayama, and N. K. Logothetis, “Magnetic resonance imaging of cortical connectivity in vivo,” Neuroimage40(2), 458–472 (2008).
[CrossRef] [PubMed]

Bria, A.

Briggman, K. L.

K. L. Briggman, M. Helmstaedter, and W. Denk, “Wiring specificity in the direction-selectivity circuit of the retina,” Nature471(7337), 183–188 (2011).
[CrossRef] [PubMed]

Callaway, E. M.

L. Luo, E. M. Callaway, and K. Svoboda, “Genetic dissection of neural circuits,” Neuron57(5), 634–660 (2008).
[CrossRef] [PubMed]

Canals, S.

S. Canals, M. Beyerlein, A. L. Keller, Y. Murayama, and N. K. Logothetis, “Magnetic resonance imaging of cortical connectivity in vivo,” Neuroimage40(2), 458–472 (2008).
[CrossRef] [PubMed]

Chen, W. R.

Chen, X.

X. Chen, U. Leischner, N. L. Rochefort, I. Nelken, and A. Konnerth, “Functional mapping of single spines in cortical neurons in vivo,” Nature475(7357), 501–505 (2011).
[CrossRef] [PubMed]

Chiovini, B.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
[CrossRef] [PubMed]

Davidson, N.

Davis, M. W.

S. Watanabe, A. Punge, G. Hollopeter, K. I. Willig, R. J. Hobson, M. W. Davis, S. W. Hell, and E. M. Jorgensen, “Protein localization in electron micrographs using fluorescence nanoscopy,” Nat. Methods8(1), 80–84 (2011).
[CrossRef] [PubMed]

Deininger, K.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Denk, W.

K. L. Briggman, M. Helmstaedter, and W. Denk, “Wiring specificity in the direction-selectivity circuit of the retina,” Nature471(7337), 183–188 (2011).
[CrossRef] [PubMed]

J. W. Lichtman and W. Denk, “The big and the small: challenges of imaging the brain’s circuits,” Science334(6056), 618–623 (2011).
[CrossRef] [PubMed]

W. Denk and H. Horstmann, “Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure,” PLoS Biol.2(11), e329 (2004).
[CrossRef] [PubMed]

Deussing, J. M.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Ding, W.

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
[CrossRef] [PubMed]

T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
[CrossRef] [PubMed]

Dodt, H. U.

K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS ONE7(3), e33916 (2012).
[CrossRef] [PubMed]

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Du, R.

Duemani Reddy, G.

G. Duemani Reddy, K. Kelleher, R. Fink, and P. Saggau, “Three-dimensional random access multiphoton microscopy for functional imaging of neuronal activity,” Nat. Neurosci.11(6), 713–720 (2008).
[CrossRef] [PubMed]

Eder, M.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Eschenko, O.

O. Eschenko, H. C. Evrard, R. M. Neves, M. Beyerlein, Y. Murayama, and N. K. Logothetis, “Tracing of noradrenergic projections using manganese-enhanced MRI,” Neuroimage59(4), 3252–3265 (2012).
[CrossRef] [PubMed]

Evrard, H. C.

O. Eschenko, H. C. Evrard, R. M. Neves, M. Beyerlein, Y. Murayama, and N. K. Logothetis, “Tracing of noradrenergic projections using manganese-enhanced MRI,” Neuroimage59(4), 3252–3265 (2012).
[CrossRef] [PubMed]

Faria, A. V.

S. Mori, K. Oishi, and A. V. Faria, “White matter atlases based on diffusion tensor imaging,” Curr. Opin. Neurol.22(4), 362–369 (2009).
[CrossRef] [PubMed]

Feng, G.

G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

Feng, Z.

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
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P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

Schaffer, C. B.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Schierloh, A.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods4(4), 331–336 (2007).
[CrossRef] [PubMed]

Schmidt, A. D.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

Schnell, S.

L. A. Harsan, D. Paul, S. Schnell, B. W. Kreher, J. Hennig, J. F. Staiger, and D. von Elverfeldt, “In vivo diffusion tensor magnetic resonance imaging and fiber tracking of the mouse brain,” NMR Biomed.23(7), 884–896 (2010).
[CrossRef] [PubMed]

Seung, H. S.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

Silvestri, L.

Smith, S. J.

K. D. Micheva and S. J. Smith, “Array tomography: a new tool for imaging the molecular architecture and ultrastructure of neural circuits,” Neuron55(1), 25–36 (2007).
[CrossRef] [PubMed]

Squier, J. A.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Staiger, J. F.

L. A. Harsan, D. Paul, S. Schnell, B. W. Kreher, J. Hennig, J. F. Staiger, and D. von Elverfeldt, “In vivo diffusion tensor magnetic resonance imaging and fiber tracking of the mouse brain,” NMR Biomed.23(7), 884–896 (2010).
[CrossRef] [PubMed]

Stelzer, E. H.

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

Sutin, J.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

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L. Luo, E. M. Callaway, and K. Svoboda, “Genetic dissection of neural circuits,” Neuron57(5), 634–660 (2008).
[CrossRef] [PubMed]

Szalay, G.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
[CrossRef] [PubMed]

Taranda, J.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

Thompson, B. D.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Tsai, P. S.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Tsien, R. Y.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Venkataraju, K. U.

T. Ragan, L. R. Kadiri, K. U. Venkataraju, K. Bahlmann, J. Sutin, J. Taranda, I. Arganda-Carreras, Y. Kim, H. S. Seung, and P. Osten, “Serial two-photon tomography for automated ex vivo mouse brain imaging,” Nat. Methods9(3), 255–258 (2012).
[CrossRef] [PubMed]

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G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
[CrossRef] [PubMed]

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G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
[CrossRef] [PubMed]

von Elverfeldt, D.

L. A. Harsan, D. Paul, S. Schnell, B. W. Kreher, J. Hennig, J. F. Staiger, and D. von Elverfeldt, “In vivo diffusion tensor magnetic resonance imaging and fiber tracking of the mouse brain,” NMR Biomed.23(7), 884–896 (2010).
[CrossRef] [PubMed]

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G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
[CrossRef] [PubMed]

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A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
[CrossRef] [PubMed]

Watanabe, S.

S. Watanabe, A. Punge, G. Hollopeter, K. I. Willig, R. J. Hobson, M. W. Davis, S. W. Hell, and E. M. Jorgensen, “Protein localization in electron micrographs using fluorescence nanoscopy,” Nat. Methods8(1), 80–84 (2011).
[CrossRef] [PubMed]

Weiler, R.

K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS ONE7(3), e33916 (2012).
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S. Watanabe, A. Punge, G. Hollopeter, K. I. Willig, R. J. Hobson, M. W. Davis, S. W. Hell, and E. M. Jorgensen, “Protein localization in electron micrographs using fluorescence nanoscopy,” Nat. Methods8(1), 80–84 (2011).
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P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
[CrossRef] [PubMed]

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H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
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A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
[CrossRef] [PubMed]

Xiong, Q.

P. S. Tsai, B. Friedman, A. I. Ifarraguerri, B. D. Thompson, V. Lev-Ram, C. B. Schaffer, Q. Xiong, R. Y. Tsien, J. A. Squier, and D. Kleinfeld, “All-optical histology using ultrashort laser pulses,” Neuron39(1), 27–41 (2003).
[CrossRef] [PubMed]

Xiong, W.

Xu, T.

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
[CrossRef] [PubMed]

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H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
[CrossRef] [PubMed]

A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
[CrossRef] [PubMed]

Yang, Z.

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
[CrossRef] [PubMed]

T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
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Zeng, S.

T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
[CrossRef] [PubMed]

H. Gong, S. Zeng, C. Yan, X. Lv, Z. Yang, T. Xu, Z. Feng, W. Ding, X. Qi, A. Li, J. Wu, and Q. Luo, “Continuously tracing brain-wide long-distance axonal projections in mice at a one-micron voxel resolution,” Neuroimage74, 87–98 (2013), doi:.
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X. Liu, T. Quan, S. Zeng, and X. Lv, “Identification of the direction of the neural network activation with a cellular resolution by fast two-photon imaging,” J. Biomed. Opt.16(8), 080506 (2011).
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A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
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D. Li, S. Zeng, X. Lv, J. Liu, R. Du, R. Jiang, W. R. Chen, and Q. Luo, “Dispersion characteristics of acousto-optic deflector for scanning Gaussian laser beam of femtosecond pulses,” Opt. Express15(8), 4726–4734 (2007).
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A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
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E. L. Bearer, X. Zhang, and R. E. Jacobs, “Live imaging of neuronal connections by magnetic resonance: Robust transport in the hippocampal-septal memory circuit in a mouse model of Down syndrome,” Neuroimage37(1), 230–242 (2007).
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T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
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T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
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[CrossRef] [PubMed]

P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010).
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G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, “Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes,” Nat. Methods9(2), 201–208 (2012).
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S. Watanabe, A. Punge, G. Hollopeter, K. I. Willig, R. J. Hobson, M. W. Davis, S. W. Hell, and E. M. Jorgensen, “Protein localization in electron micrographs using fluorescence nanoscopy,” Nat. Methods8(1), 80–84 (2011).
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E. L. Bearer, X. Zhang, and R. E. Jacobs, “Live imaging of neuronal connections by magnetic resonance: Robust transport in the hippocampal-septal memory circuit in a mouse model of Down syndrome,” Neuroimage37(1), 230–242 (2007).
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G. Feng, R. H. Mellor, M. Bernstein, C. Keller-Peck, Q. T. Nguyen, M. Wallace, J. M. Nerbonne, J. W. Lichtman, and J. R. Sanes, “Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP,” Neuron28(1), 41–51 (2000).
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Sci Rep (1)

T. Quan, T. Zheng, Z. Yang, W. Ding, S. Li, J. Li, H. Zhou, Q. Luo, H. Gong, and S. Zeng, “NeuroGPS: automated localization of neurons for brain circuits using L1 minimization model,” Sci Rep3, 1414 (2013), doi:.
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Science (3)

A. Li, H. Gong, B. Zhang, Q. Wang, C. Yan, J. Wu, Q. Liu, S. Zeng, and Q. Luo, “Micro-optical sectioning tomography to obtain a high-resolution atlas of the mouse brain,” Science330(6009), 1404–1408 (2010).
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Supplementary Material (4)

» Media 1: AVI (1054 KB)     
» Media 2: AVI (624 KB)     
» Media 3: AVI (1054 KB)     
» Media 4: AVI (624 KB)     

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Figures (6)

Fig. 1
Fig. 1

Optical scheme of 2P-fMOST system. The whole 2p-MOST system was composed of (a) dispersion compensation, (b) fast scanning, (c) microscope, and (d) motion and sectioning modules. The relay optics consists of scan lens (SL, 150 mm) and tube lens (TL, 180 mm) images of the AOD on the back aperture of the objective. (e) Lateral normal fluorescence intensity distribution of the bead, the full width at half maximum (FWHM) value is approximately 0.45 µm. (f) Axial normal fluorescence intensity distribution of the bead, FWHM value is approximately 1.68 μm.

Fig. 2
Fig. 2

Generation and compensation of astigmatism of 2p-fMOST system (a) AOD was driven with chirp acoustic frequency; the light spot lengthens in the y-direction after AOD. With the compensation of the converged cylindrical lens, the light spot becomes recollimated. (b) Image of pollen without astigmatism compensation. (c) Image of pollen with astigmatism compensation. Scale bar is 10 µm in (b) and (c).

Fig. 3
Fig. 3

Imaging strategy of 2p-fMOST system. (a) The blurred area represents the area that has not been imaged, the laser scanning in the y-direction is shown by the red line, and the scanning in the x-direction was accomplished by the stage. (b) Different imaging areas were set according to brain profile. The red frame stands for the real imaging area, and the green areas depict the shape of brain in the different z-planes.

Fig. 4
Fig. 4

Imaging performance on the entire brain. (a) 3D volume presentation of an EGFP mouse brain. The superimposed plane refers to coronal sections shown in (b). (b) 100 µm z-plane maximum projection image, coronal view, scale bar: 1 mm. (c) Enlarged view of the neurons signed as red frame in (b), contrast was enhanced 30% for a better visibility, scale bar: 200 µm. (d) Enlarged view signed as red frame in (c), scale bar: 30 µm. (e) Enlarged view of dendrites signed as red frame in (d), scale bar: 5 µm. dendritic spines are as white arrows pointed. (f) Enlarged view of the lower frame in (c), scale bar: 5 µm, axon is as white arrows pointed (See Media 1).

Fig. 5
Fig. 5

No distortion in imaging after sectioning. (a) A 60 µm z-plane maximum projection image of neurons, depth from 60 μm to 120 μm below the surface of sample. (b) A 60 µm z-plane maximum projection image of neurons with z-depth from 30 μm to 90 μm below the surface after a 30 μm thick sectioning. (c) Merged image of (a) and (b), the mapped areas of (a) and (b) shown in yellow. (d), (e), and (f) are the enlarged views of the white frame in (a), (b), and (c) respectively. High superposition in any typical structure was shown, including soma and slim dendrites as white arrows pointed. Scale bar is 1 mm in (a), (b), and (c), and 20 μm in (d), (e), and (f).

Fig. 6
Fig. 6

Tracing long-range axonal projections in the whole brain. (a) 8 axonal projections were traced in 3D, different colors represent different axonal projections. (b) Coronal view of the projections. (c) Sagittal views of the projections. (d) Horizon view of the projections. Scale bar is 1 mm in (a) to (d) (See Media 2).

Equations (1)

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f= v 2 λα .

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