Simultaneous two-photon activation and imaging of neural activity based on spectral–temporal modulation of supercontinuum light
| Autor: | Ronit Barkalifa, Eric J. Chaney, Catherine A. Christian-Hinman, Yuan-Zhi Liu, Rishyashring R. Iyer, Daniel A. Llano, Connor D. Courtney, Stephen A. Boppart, Sixian You, Baher A. Ibrahim, Carlos Renteria, Haohua Tu, Mantas Zurauskas |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Paper
Materials science two-photon fluorescence microscopy Neuroscience (miscellaneous) Physics::Optics neurons 01 natural sciences two-photon optogenetics law.invention 010309 optics 03 medical and health sciences 0302 clinical medicine Optics Two-photon excitation microscopy law 0103 physical sciences Radiology Nuclear Medicine and imaging Quantitative Biology::Neurons and Cognition Radiological and Ultrasound Technology business.industry Nonlinear optics Laser Research Papers Supercontinuum calcium imaging supercontinuum Femtosecond business photonic crystal fiber Ultrashort pulse 030217 neurology & neurosurgery Excitation Photonic-crystal fiber |
| Zdroj: | Neurophotonics |
| ISSN: | 2329-4248 2329-423X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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