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Simultaneous two-photon activation and imaging of neural activity based on spectral-temporal modulation of supercontinuum light.
Neurophotonics ( IF 5.3 ) Pub Date : 2020-11-03 , DOI: 10.1117/1.nph.7.4.045007
Yuan-Zhi Liu 1, 2 , Carlos Renteria 1, 3 , Connor D Courtney 1, 4 , Baher Ibrahim 1 , Sixian You 1, 2, 5 , Eric J Chaney 1 , Ronit Barkalifa 1 , Rishyashring R Iyer 1, 2 , Mantas Zurauskas 1 , Haohua Tu 1 , Daniel A Llano 1, 4, 6 , Catherine A Christian-Hinman 1, 4, 6 , Stephen A Boppart 1, 2, 3, 4, 5, 7
Affiliation  

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.

中文翻译:

基于超连续光的光谱-时间调制,同时进行双光子激活和神经活动成像。

意义神经科学中非线性光学的最新进展集中在使用两个超快激光进行活动成像和光遗传学刺激上。宽带飞秒光源可通过光谱分离实现彩色目标激发,从而消除了对多台激光器的需求。目的我们提出一种基于光子晶体光纤(PCF)的超连续谱源,分别用于光谱分辨双光子(2P)成像和GCaMP6s和C1V1-mCherry的激发。方法使用20 MHz重复频率飞秒激光泵浦PCF,以产生超连续光,该光被光谱分离,压缩并重组为图像GCaMP6(930 nm激发),并刺激光遗传蛋白C1V1-mCherry(1060 nm)。励磁)。电流螺旋扫描在单细胞水平上用于多光子激发,高速共振扫描用于钙活性成像。结果使用连续波激光验证光遗传学激活的功能,然后进行定向2P激发。这些实验的结果表明,超连续谱光源可用于同时进行的单细胞激发和钙成像。结论基于PCF的超连续谱光源可用于同时成像和激发脑组织中钙动力学。抽运的PCF可以用作强大的光源,用于成像和激活神经活动,并克服了与多激光方法相关的有限光谱和空间。结果使用连续波激光验证光遗传学激活的功能,然后进行定向2P激发。这些实验的结果表明,超连续谱光源可用于同时进行的单细胞激发和钙成像。结论基于PCF的超连续谱光源可用于同时成像和激发脑组织中钙动力学。抽运的PCF可以用作强大的光源,用于成像和激活神经活动,并克服了与多激光方法相关的有限光谱和空间。结果使用连续波激光验证光遗传学激活的功能,然后进行定向2P激发。这些实验的结果表明,超连续谱光源可用于同时进行的单细胞激发和钙成像。结论基于PCF的超连续谱光源可用于同时成像和激发脑组织中钙动力学。抽运的PCF可以用作强大的光源,用于成像和激活神经活动,并克服了与多激光方法相关的有限光谱和空间。结论基于PCF的超连续谱光源可用于同时成像和激发脑组织中钙动力学。抽运的PCF可以用作强大的光源,用于成像和激活神经活动,并克服了与多激光方法相关的有限光谱和空间。结论基于PCF的超连续谱光源可用于同时成像和激发脑组织中钙动力学。抽运的PCF可以用作强大的光源,用于成像和激活神经活动,并克服了与多激光方法相关的有限光谱和空间。
更新日期:2020-11-03
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