Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution

Tonmoy Chakraborty; Meghan K. Driscoll; Elise Jeffery; Malea M. Murphy; Philippe Roudot; Bo Jui Chang; Saumya Vora; Wen Mai Wong; Cara D. Nielson; Hua Zhang; Vladimir Zhemkov; Chitkale Hiremath; Estanislao Daniel De La Cruz; Yating Yi; Ilya Bezprozvanny; Hu Zhao; Raju Tomer; Rainer Heintzmann; Julian Meeks; Denise K. Marciano; Sean J. Morrison; Gaudenz Danuser; Kevin M. Dean; Reto P Fiolka

doi:10.1038/s41592-019-0615-4

Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution

Tonmoy Chakraborty, Meghan K. Driscoll, Elise Jeffery, Malea M. Murphy, Philippe Roudot, Bo Jui Chang, Saumya Vora, Wen Mai Wong, Cara D. Nielson, Hua Zhang, Vladimir Zhemkov, Chitkale Hiremath, Estanislao Daniel De La Cruz, Yating Yi, Ilya Bezprozvanny, Hu Zhao, Raju Tomer, Rainer Heintzmann, Julian Meeks, Denise K. MarcianoSean J. Morrison, Gaudenz Danuser, Kevin M. Dean, Reto P Fiolka

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73 Scopus citations

Abstract

We present cleared-tissue axially swept light-sheet microscopy (ctASLM), which enables isotropic, subcellular resolution imaging with high optical sectioning capability and a large field of view over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and non-aqueous chemically cleared tissue preparations. Depending on the optical configuration, ctASLM provides up to 260 nm of axial resolution, a three to tenfold improvement over confocal and other reported cleared-tissue light-sheet microscopes. We imaged millimeter-scale cleared tissues with subcellular three-dimensional resolution, which enabled automated detection of multicellular tissue architectures, individual cells, synaptic spines and rare cell–cell interactions.

Original language	English (US)
Pages (from-to)	1109-1113
Number of pages	5
Journal	Nature methods
Volume	16
Issue number	11
DOIs	https://doi.org/10.1038/s41592-019-0615-4
State	Published - Nov 1 2019

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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Chakraborty, T., Driscoll, M. K., Jeffery, E., Murphy, M. M., Roudot, P., Chang, B. J., Vora, S., Wong, W. M., Nielson, C. D., Zhang, H., Zhemkov, V., Hiremath, C., De La Cruz, E. D., Yi, Y., Bezprozvanny, I., Zhao, H., Tomer, R., Heintzmann, R., Meeks, J., ... Fiolka, R. P. (2019). Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution. Nature methods, 16(11), 1109-1113. https://doi.org/10.1038/s41592-019-0615-4

Chakraborty, T, Driscoll, MK, Jeffery, E, Murphy, MM, Roudot, P, Chang, BJ, Vora, S, Wong, WM, Nielson, CD, Zhang, H, Zhemkov, V, Hiremath, C, De La Cruz, ED, Yi, Y, Bezprozvanny, I, Zhao, H, Tomer, R, Heintzmann, R, Meeks, J, Marciano, DK , Morrison, SJ , Danuser, G , Dean, KM & Fiolka, RP 2019, 'Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution', Nature methods, vol. 16, no. 11, pp. 1109-1113. https://doi.org/10.1038/s41592-019-0615-4

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title = "Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution",

abstract = "We present cleared-tissue axially swept light-sheet microscopy (ctASLM), which enables isotropic, subcellular resolution imaging with high optical sectioning capability and a large field of view over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and non-aqueous chemically cleared tissue preparations. Depending on the optical configuration, ctASLM provides up to 260 nm of axial resolution, a three to tenfold improvement over confocal and other reported cleared-tissue light-sheet microscopes. We imaged millimeter-scale cleared tissues with subcellular three-dimensional resolution, which enabled automated detection of multicellular tissue architectures, individual cells, synaptic spines and rare cell–cell interactions.",

author = "Tonmoy Chakraborty and Driscoll, {Meghan K.} and Elise Jeffery and Murphy, {Malea M.} and Philippe Roudot and Chang, {Bo Jui} and Saumya Vora and Wong, {Wen Mai} and Nielson, {Cara D.} and Hua Zhang and Vladimir Zhemkov and Chitkale Hiremath and {De La Cruz}, {Estanislao Daniel} and Yating Yi and Ilya Bezprozvanny and Hu Zhao and Raju Tomer and Rainer Heintzmann and Julian Meeks and Marciano, {Denise K.} and Morrison, {Sean J.} and Gaudenz Danuser and Dean, {Kevin M.} and Fiolka, {Reto P}",

note = "Funding Information: We thank the Cancer Prevention Research Institute of Texas (grant nos. RR160057 to R.F. and R1225 to G.D.) for their generous funding, as well as the National Institutes of Health (grant nos. F32GM116370 and K99GM123221 to M.K.D., R01GM067230 to G.D., R01AG055577 and R01NS056224 to I.B., R01DK118032 and R01DK099478 to D.M., R01DC015784 and R21NS104826 to J.M., R33CA235254 and R35GM133522 to R.F., and DP2MH119423 to R.T.). R.H. acknowledges support by the Collaborative Research Center SFB 1278 (PolyTarget, project C04) funded by the Deutsche Forschungsgemeinschaft. We are grateful to the Live cell imaging facility at UT Southwestern for access to the Zeiss Airyscan confocal microscope (supported by grant no. 1S10OD021684-01). We thank D. Saucier and J. Amatruda for providing the zebrafish specimens and E. Sapoznik for his assistance with computer-aided design. Publisher Copyright: {\textcopyright} 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.",

year = "2019",

month = nov,

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doi = "10.1038/s41592-019-0615-4",

language = "English (US)",

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T1 - Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution

AU - Chakraborty, Tonmoy

AU - Driscoll, Meghan K.

AU - Jeffery, Elise

AU - Murphy, Malea M.

AU - Roudot, Philippe

AU - Chang, Bo Jui

AU - Vora, Saumya

AU - Wong, Wen Mai

AU - Nielson, Cara D.

AU - Zhang, Hua

AU - Zhemkov, Vladimir

AU - Hiremath, Chitkale

AU - De La Cruz, Estanislao Daniel

AU - Yi, Yating

AU - Bezprozvanny, Ilya

AU - Zhao, Hu

AU - Tomer, Raju

AU - Heintzmann, Rainer

AU - Meeks, Julian

AU - Marciano, Denise K.

AU - Morrison, Sean J.

AU - Danuser, Gaudenz

AU - Dean, Kevin M.

AU - Fiolka, Reto P

N1 - Funding Information: We thank the Cancer Prevention Research Institute of Texas (grant nos. RR160057 to R.F. and R1225 to G.D.) for their generous funding, as well as the National Institutes of Health (grant nos. F32GM116370 and K99GM123221 to M.K.D., R01GM067230 to G.D., R01AG055577 and R01NS056224 to I.B., R01DK118032 and R01DK099478 to D.M., R01DC015784 and R21NS104826 to J.M., R33CA235254 and R35GM133522 to R.F., and DP2MH119423 to R.T.). R.H. acknowledges support by the Collaborative Research Center SFB 1278 (PolyTarget, project C04) funded by the Deutsche Forschungsgemeinschaft. We are grateful to the Live cell imaging facility at UT Southwestern for access to the Zeiss Airyscan confocal microscope (supported by grant no. 1S10OD021684-01). We thank D. Saucier and J. Amatruda for providing the zebrafish specimens and E. Sapoznik for his assistance with computer-aided design. Publisher Copyright: © 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.

PY - 2019/11/1

Y1 - 2019/11/1

N2 - We present cleared-tissue axially swept light-sheet microscopy (ctASLM), which enables isotropic, subcellular resolution imaging with high optical sectioning capability and a large field of view over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and non-aqueous chemically cleared tissue preparations. Depending on the optical configuration, ctASLM provides up to 260 nm of axial resolution, a three to tenfold improvement over confocal and other reported cleared-tissue light-sheet microscopes. We imaged millimeter-scale cleared tissues with subcellular three-dimensional resolution, which enabled automated detection of multicellular tissue architectures, individual cells, synaptic spines and rare cell–cell interactions.

AB - We present cleared-tissue axially swept light-sheet microscopy (ctASLM), which enables isotropic, subcellular resolution imaging with high optical sectioning capability and a large field of view over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and non-aqueous chemically cleared tissue preparations. Depending on the optical configuration, ctASLM provides up to 260 nm of axial resolution, a three to tenfold improvement over confocal and other reported cleared-tissue light-sheet microscopes. We imaged millimeter-scale cleared tissues with subcellular three-dimensional resolution, which enabled automated detection of multicellular tissue architectures, individual cells, synaptic spines and rare cell–cell interactions.

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AN - SCOPUS:85074283217

VL - 16

SP - 1109

EP - 1113

JO - Nature Methods

JF - Nature Methods

SN - 1548-7091

IS - 11

ER -

Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution

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