Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light

Yuan-Zhi Liu; Carlos Renteria; Connor D Courtney; Baher Ibrahim; Sixian You; Eric J Chaney; Ronit Barkalifa; Rishyashring R Iyer; Mantas Zurauskas; Haohua Tu; Daniel A Llano; Catherine A Christian-Hinman; Stephen A Boppart

doi:10.1117/1.NPh.7.4.045007

Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light

Yuan-Zhi Liu, Carlos Renteria, Connor D Courtney, Baher Ibrahim, Sixian You, Eric J Chaney, Ronit Barkalifa, Rishyashring R Iyer, Mantas Zurauskas, Haohua Tu, Daniel A Llano, Catherine A Christian-Hinman, Stephen A Boppart

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

Abstract

Significance: Recent advances in nonlinear optics in neuroscience have focused on using two ultrafast lasers for activity imaging and optogenetic stimulation. Broadband femtosecond light sources can obviate the need for multiple lasers by spectral separation for chromatically targeted excitation.

Aim: We present a photonic crystal fiber (PCF)-based supercontinuum source for spectrally resolved two-photon (2P) imaging and excitation of GCaMP6s and C1V1-mCherry, respectively.

Approach: A PCF is pumped using a 20-MHz repetition rate femtosecond laser to generate a supercontinuum of light, which is spectrally separated, compressed, and recombined to image GCaMP6s (930 nm excitation) and stimulate the optogenetic protein, C1V1-mCherry (1060 nm excitation). Galvanometric spiral scanning is employed on a single-cell level for multiphoton excitation and high-speed resonant scanning is employed for imaging of calcium activity.

Results: Continuous wave lasers were used to verify functionality of optogenetic activation followed by directed 2P excitation. Results from these experiments demonstrate the utility of a supercontinuum light source for simultaneous, single-cell excitation and calcium imaging.

Conclusions: A PCF-based supercontinuum light source was employed for simultaneous imaging and excitation of calcium dynamics in brain tissue. Pumped PCFs can serve as powerful light sources for imaging and activation of neural activity, and overcome the limited spectra and space associated with multilaser approaches.

Original language	English (US)
Pages (from-to)	045007
Journal	Neurophotonics
Volume	7
Issue number	4
DOIs	https://doi.org/10.1117/1.NPh.7.4.045007
State	Published - Oct 2020

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Liu, Y.-Z., Renteria, C., Courtney, C. D., Ibrahim, B., You, S., Chaney, E. J., Barkalifa, R., Iyer, R. R., Zurauskas, M., Tu, H., Llano, D. A., Christian-Hinman, C. A., & Boppart, S. A. (2020). Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light. Neurophotonics, 7(4), 045007. https://doi.org/10.1117/1.NPh.7.4.045007

Liu, Y-Z, Renteria, C, Courtney, CD, Ibrahim, B, You, S, Chaney, EJ, Barkalifa, R, Iyer, RR, Zurauskas, M, Tu, H, Llano, DA , Christian-Hinman, CA & Boppart, SA 2020, 'Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light', Neurophotonics, vol. 7, no. 4, pp. 045007. https://doi.org/10.1117/1.NPh.7.4.045007

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title = "Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light",

abstract = "Significance: Recent advances in nonlinear optics in neuroscience have focused on using two ultrafast lasers for activity imaging and optogenetic stimulation. Broadband femtosecond light sources can obviate the need for multiple lasers by spectral separation for chromatically targeted excitation.Aim: We present a photonic crystal fiber (PCF)-based supercontinuum source for spectrally resolved two-photon (2P) imaging and excitation of GCaMP6s and C1V1-mCherry, respectively.Approach: A PCF is pumped using a 20-MHz repetition rate femtosecond laser to generate a supercontinuum of light, which is spectrally separated, compressed, and recombined to image GCaMP6s (930 nm excitation) and stimulate the optogenetic protein, C1V1-mCherry (1060 nm excitation). Galvanometric spiral scanning is employed on a single-cell level for multiphoton excitation and high-speed resonant scanning is employed for imaging of calcium activity.Results: Continuous wave lasers were used to verify functionality of optogenetic activation followed by directed 2P excitation. Results from these experiments demonstrate the utility of a supercontinuum light source for simultaneous, single-cell excitation and calcium imaging.Conclusions: A PCF-based supercontinuum light source was employed for simultaneous imaging and excitation of calcium dynamics in brain tissue. Pumped PCFs can serve as powerful light sources for imaging and activation of neural activity, and overcome the limited spectra and space associated with multilaser approaches.",

author = "Yuan-Zhi Liu and Carlos Renteria and Courtney, {Connor D} and Baher Ibrahim and Sixian You and Chaney, {Eric J} and Ronit Barkalifa and Iyer, {Rishyashring R} and Mantas Zurauskas and Haohua Tu and Llano, {Daniel A} and Christian-Hinman, {Catherine A} and Boppart, {Stephen A}",

note = "{\textcopyright} The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.",

year = "2020",

month = oct,

doi = "10.1117/1.NPh.7.4.045007",

language = "English (US)",

volume = "7",

pages = "045007",

journal = "Neurophotonics",

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TY - JOUR

T1 - Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light

AU - Liu, Yuan-Zhi

AU - Renteria, Carlos

AU - Courtney, Connor D

AU - Ibrahim, Baher

AU - You, Sixian

AU - Chaney, Eric J

AU - Barkalifa, Ronit

AU - Iyer, Rishyashring R

AU - Zurauskas, Mantas

AU - Tu, Haohua

AU - Llano, Daniel A

AU - Christian-Hinman, Catherine A

AU - Boppart, Stephen A

N1 - © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

PY - 2020/10

Y1 - 2020/10

N2 - Significance: Recent advances in nonlinear optics in neuroscience have focused on using two ultrafast lasers for activity imaging and optogenetic stimulation. Broadband femtosecond light sources can obviate the need for multiple lasers by spectral separation for chromatically targeted excitation.Aim: We present a photonic crystal fiber (PCF)-based supercontinuum source for spectrally resolved two-photon (2P) imaging and excitation of GCaMP6s and C1V1-mCherry, respectively.Approach: A PCF is pumped using a 20-MHz repetition rate femtosecond laser to generate a supercontinuum of light, which is spectrally separated, compressed, and recombined to image GCaMP6s (930 nm excitation) and stimulate the optogenetic protein, C1V1-mCherry (1060 nm excitation). Galvanometric spiral scanning is employed on a single-cell level for multiphoton excitation and high-speed resonant scanning is employed for imaging of calcium activity.Results: Continuous wave lasers were used to verify functionality of optogenetic activation followed by directed 2P excitation. Results from these experiments demonstrate the utility of a supercontinuum light source for simultaneous, single-cell excitation and calcium imaging.Conclusions: A PCF-based supercontinuum light source was employed for simultaneous imaging and excitation of calcium dynamics in brain tissue. Pumped PCFs can serve as powerful light sources for imaging and activation of neural activity, and overcome the limited spectra and space associated with multilaser approaches.

AB - Significance: Recent advances in nonlinear optics in neuroscience have focused on using two ultrafast lasers for activity imaging and optogenetic stimulation. Broadband femtosecond light sources can obviate the need for multiple lasers by spectral separation for chromatically targeted excitation.Aim: We present a photonic crystal fiber (PCF)-based supercontinuum source for spectrally resolved two-photon (2P) imaging and excitation of GCaMP6s and C1V1-mCherry, respectively.Approach: A PCF is pumped using a 20-MHz repetition rate femtosecond laser to generate a supercontinuum of light, which is spectrally separated, compressed, and recombined to image GCaMP6s (930 nm excitation) and stimulate the optogenetic protein, C1V1-mCherry (1060 nm excitation). Galvanometric spiral scanning is employed on a single-cell level for multiphoton excitation and high-speed resonant scanning is employed for imaging of calcium activity.Results: Continuous wave lasers were used to verify functionality of optogenetic activation followed by directed 2P excitation. Results from these experiments demonstrate the utility of a supercontinuum light source for simultaneous, single-cell excitation and calcium imaging.Conclusions: A PCF-based supercontinuum light source was employed for simultaneous imaging and excitation of calcium dynamics in brain tissue. Pumped PCFs can serve as powerful light sources for imaging and activation of neural activity, and overcome the limited spectra and space associated with multilaser approaches.

U2 - 10.1117/1.NPh.7.4.045007

DO - 10.1117/1.NPh.7.4.045007

M3 - Article

C2 - 33163545

SN - 2329-423X

VL - 7

SP - 045007

JO - Neurophotonics

JF - Neurophotonics

IS - 4

ER -

Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light

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