A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice

Nicolò Accanto; François G.C. Blot; Antonio Lorca-Cámara; Valeria Zampini; Florence Bui; Christophe Tourain; Noam Badt; Ori Katz; Valentina Emiliani

doi:10.1016/j.neuron.2022.10.030

A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice

Nicolò Accanto^*, François G.C. Blot, Antonio Lorca-Cámara, Valeria Zampini, Florence Bui, Christophe Tourain, Noam Badt, Ori Katz, Valentina Emiliani^*

^*Corresponding author for this work

Department of Applied Physics

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

We developed a flexible two-photon microendoscope (2P-FENDO) capable of all-optical brain investigation at near cellular resolution in freely moving mice. The system performs fast two-photon (2P) functional imaging and 2P holographic photostimulation of single and multiple cells using axially confined extended spots. Proof-of-principle experiments were performed in freely moving mice co-expressing jGCaMP7s and the opsin ChRmine in the visual or barrel cortex. On a field of view of 250 μm in diameter, we demonstrated functional imaging at a frame rate of up to 50 Hz and precise photostimulation of selected groups of cells. With the capability to simultaneously image and control defined neuronal networks in freely moving animals, 2P-FENDO will enable a precise investigation of neuronal functions in the brain during naturalistic behaviors.

Original language	American English
Pages (from-to)	176-189.e6
Journal	Neuron
Volume	111
Issue number	2
DOIs	https://doi.org/10.1016/j.neuron.2022.10.030
State	Published - 18 Jan 2023

Bibliographical note

Funding Information:
We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000 ), the Fondation Bettencourt Schueller (Prix Coups d’élan pour la recherche française), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01 ), the Human Frontiers Science Program (grant RGP0015/2016 ), Imagine Nano, the ERC Advanced Grant HOLOVIS ( ERC-2019-AdG ; award no. 885090 ), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787 ), the Israel Science Foundation ( 1361/18 ), and the Israeli Ministry of Science and Technology (grant 712845 ). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omnès (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation.

Funding Information:
We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000), the Fondation Bettencourt Schueller (Prix Coups d’élan pour la recherche française), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01), the Human Frontiers Science Program (grant RGP0015/2016), Imagine Nano, the ERC Advanced Grant HOLOVIS (ERC-2019-AdG; award no. 885090), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787), the Israel Science Foundation (1361/18), and the Israeli Ministry of Science and Technology (grant 712845). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omnès (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation. N.A. A.L.-C. and C.T. built the optical setup. N.A. and A.L.-C. developed the acquisition software and performed the optical characterization. O.K. conceived the experiment for measuring the ICDD. N.B. measured and analyzed the ICDD. F.G.C.B. and C.T. designed and developed the implant for freely moving experiments. F.G.C.B. V.Z. and F.B. developed surgical procedures and performed viral injections. V.Z. and F.B. acquired in vivo and in vitro z stacks and preliminary calcium imaging data in vivo. F.G.C.B. and A.L.-C. performed imaging and photostimulation experiments in freely moving mice. N.A. F.G.C.B. and A.L.-C. analyzed the data. N.A. F.G.C.B. and V.E. wrote the paper with the contribution of all the authors. N.A. and V.E. conceived and supervised the project. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Publisher Copyright:
© 2022 Elsevier Inc.

Keywords

2-photon calcium imaging
2-photon optogenetic photostimulation
GRIN lenses
all-optical neuronal circuit manipulation
computer-generated holography
freely moving mice
microendoscopy
optical fiber bundles

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10.1016/j.neuron.2022.10.030

Cite this

@article{773059b7493b4a9f9ad24be62e63406a,

title = "A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice",

abstract = "We developed a flexible two-photon microendoscope (2P-FENDO) capable of all-optical brain investigation at near cellular resolution in freely moving mice. The system performs fast two-photon (2P) functional imaging and 2P holographic photostimulation of single and multiple cells using axially confined extended spots. Proof-of-principle experiments were performed in freely moving mice co-expressing jGCaMP7s and the opsin ChRmine in the visual or barrel cortex. On a field of view of 250 μm in diameter, we demonstrated functional imaging at a frame rate of up to 50 Hz and precise photostimulation of selected groups of cells. With the capability to simultaneously image and control defined neuronal networks in freely moving animals, 2P-FENDO will enable a precise investigation of neuronal functions in the brain during naturalistic behaviors.",

keywords = "2-photon calcium imaging, 2-photon optogenetic photostimulation, GRIN lenses, all-optical neuronal circuit manipulation, computer-generated holography, freely moving mice, microendoscopy, optical fiber bundles",

author = "Nicol{\`o} Accanto and Blot, {Fran{\c c}ois G.C.} and Antonio Lorca-C{\'a}mara and Valeria Zampini and Florence Bui and Christophe Tourain and Noam Badt and Ori Katz and Valentina Emiliani",

note = "Funding Information: We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000 ), the Fondation Bettencourt Schueller (Prix Coups d{\textquoteright}{\'e}lan pour la recherche fran{\c c}aise), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01 ), the Human Frontiers Science Program (grant RGP0015/2016 ), Imagine Nano, the ERC Advanced Grant HOLOVIS ( ERC-2019-AdG ; award no. 885090 ), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787 ), the Israel Science Foundation ( 1361/18 ), and the Israeli Ministry of Science and Technology (grant 712845 ). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omn{\`e}s (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation. Funding Information: We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000), the Fondation Bettencourt Schueller (Prix Coups d{\textquoteright}{\'e}lan pour la recherche fran{\c c}aise), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01), the Human Frontiers Science Program (grant RGP0015/2016), Imagine Nano, the ERC Advanced Grant HOLOVIS (ERC-2019-AdG; award no. 885090), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787), the Israel Science Foundation (1361/18), and the Israeli Ministry of Science and Technology (grant 712845). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omn{\`e}s (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation. N.A. A.L.-C. and C.T. built the optical setup. N.A. and A.L.-C. developed the acquisition software and performed the optical characterization. O.K. conceived the experiment for measuring the ICDD. N.B. measured and analyzed the ICDD. F.G.C.B. and C.T. designed and developed the implant for freely moving experiments. F.G.C.B. V.Z. and F.B. developed surgical procedures and performed viral injections. V.Z. and F.B. acquired in vivo and in vitro z stacks and preliminary calcium imaging data in vivo. F.G.C.B. and A.L.-C. performed imaging and photostimulation experiments in freely moving mice. N.A. F.G.C.B. and A.L.-C. analyzed the data. N.A. F.G.C.B. and V.E. wrote the paper with the contribution of all the authors. N.A. and V.E. conceived and supervised the project. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",

year = "2023",

month = jan,

day = "18",

doi = "10.1016/j.neuron.2022.10.030",

language = "American English",

volume = "111",

pages = "176--189.e6",

journal = "Neuron",

issn = "0896-6273",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice

AU - Accanto, Nicolò

AU - Blot, François G.C.

AU - Lorca-Cámara, Antonio

AU - Zampini, Valeria

AU - Bui, Florence

AU - Tourain, Christophe

AU - Badt, Noam

AU - Katz, Ori

AU - Emiliani, Valentina

N1 - Funding Information: We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000 ), the Fondation Bettencourt Schueller (Prix Coups d’élan pour la recherche française), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01 ), the Human Frontiers Science Program (grant RGP0015/2016 ), Imagine Nano, the ERC Advanced Grant HOLOVIS ( ERC-2019-AdG ; award no. 885090 ), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787 ), the Israel Science Foundation ( 1361/18 ), and the Israeli Ministry of Science and Technology (grant 712845 ). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omnès (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation. Funding Information: We acknowledge support from the IHU FOReSIGHT grant (grant P-ALLOP3-IHU-000), the Fondation Bettencourt Schueller (Prix Coups d’élan pour la recherche française), the Axa research funding, the Agence National de la Recherche (grant ANR 2MEnHoloMD - 19-CE19-0001-01), the Human Frontiers Science Program (grant RGP0015/2016), Imagine Nano, the ERC Advanced Grant HOLOVIS (ERC-2019-AdG; award no. 885090), the ERC Horizon 2020 H2020-ICT (DEEPER, 101016787), the Israel Science Foundation (1361/18), and the Israeli Ministry of Science and Technology (grant 712845). We would like to thank Serge Charpak, Marine Tournissac, and Manon Omnès (Vision Institute, Paris) for their helpful discussions and pilot imaging experiments; Andrea Giovannucci for the help with CaImAn; Aaron Benson Wong (ErasmusMC Rotterdam) for the help with the calcium imaging data analysis; Vincent de Sars for the WaveFront IV software development; and Manuel Simonutti, Julie Degardin, and Quenol Cesar from the animal facility of the Institut de la vision for their help and support with the animal experimentation. N.A. A.L.-C. and C.T. built the optical setup. N.A. and A.L.-C. developed the acquisition software and performed the optical characterization. O.K. conceived the experiment for measuring the ICDD. N.B. measured and analyzed the ICDD. F.G.C.B. and C.T. designed and developed the implant for freely moving experiments. F.G.C.B. V.Z. and F.B. developed surgical procedures and performed viral injections. V.Z. and F.B. acquired in vivo and in vitro z stacks and preliminary calcium imaging data in vivo. F.G.C.B. and A.L.-C. performed imaging and photostimulation experiments in freely moving mice. N.A. F.G.C.B. and A.L.-C. analyzed the data. N.A. F.G.C.B. and V.E. wrote the paper with the contribution of all the authors. N.A. and V.E. conceived and supervised the project. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Publisher Copyright: © 2022 Elsevier Inc.

PY - 2023/1/18

Y1 - 2023/1/18

N2 - We developed a flexible two-photon microendoscope (2P-FENDO) capable of all-optical brain investigation at near cellular resolution in freely moving mice. The system performs fast two-photon (2P) functional imaging and 2P holographic photostimulation of single and multiple cells using axially confined extended spots. Proof-of-principle experiments were performed in freely moving mice co-expressing jGCaMP7s and the opsin ChRmine in the visual or barrel cortex. On a field of view of 250 μm in diameter, we demonstrated functional imaging at a frame rate of up to 50 Hz and precise photostimulation of selected groups of cells. With the capability to simultaneously image and control defined neuronal networks in freely moving animals, 2P-FENDO will enable a precise investigation of neuronal functions in the brain during naturalistic behaviors.

AB - We developed a flexible two-photon microendoscope (2P-FENDO) capable of all-optical brain investigation at near cellular resolution in freely moving mice. The system performs fast two-photon (2P) functional imaging and 2P holographic photostimulation of single and multiple cells using axially confined extended spots. Proof-of-principle experiments were performed in freely moving mice co-expressing jGCaMP7s and the opsin ChRmine in the visual or barrel cortex. On a field of view of 250 μm in diameter, we demonstrated functional imaging at a frame rate of up to 50 Hz and precise photostimulation of selected groups of cells. With the capability to simultaneously image and control defined neuronal networks in freely moving animals, 2P-FENDO will enable a precise investigation of neuronal functions in the brain during naturalistic behaviors.

KW - 2-photon calcium imaging

KW - 2-photon optogenetic photostimulation

KW - GRIN lenses

KW - all-optical neuronal circuit manipulation

KW - computer-generated holography

KW - freely moving mice

KW - microendoscopy

KW - optical fiber bundles

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U2 - 10.1016/j.neuron.2022.10.030

DO - 10.1016/j.neuron.2022.10.030

M3 - Article

C2 - 36395773

AN - SCOPUS:85146087849

SN - 0896-6273

VL - 111

SP - 176-189.e6

JO - Neuron

JF - Neuron

IS - 2

ER -

A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice

Abstract

Bibliographical note

Keywords

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