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：

---

基于超连续谱光的光谱时间调制的神经活动的同时双光子激活和成像。

意义神经科学非线性光学的最新进展集中在使用两个超快激光器进行活动成像和光遗传学刺激。宽带飞秒光源可以通过光谱分离进行色彩目标激发，从而消除对多个激光器的需求。目的 我们提出了一种基于光子晶体光纤 (PCF) 的超连续谱源，分别用于 GCaMP6s 和 C1V1-mCherry 的光谱分辨双光子 (2P) 成像和激发。方法 使用 20 MHz 重复率飞秒激光器泵浦 PCF，产生超连续光，对光进行光谱分离、压缩和重组，以对 GCaMP6s 进行成像（930 nm 激发）并刺激光遗传学蛋白 C1V1-mCherry（1060 nm）励磁）。在单细胞水平上采用检流螺旋扫描进行多光子激发，并采用高速共振扫描对钙活性进行成像。结果 使用连续波激光器来验证光遗传学激活和定向 2P 激发的功能。这些实验的结果证明了超连续谱光源在同步单细胞激发和钙成像中的实用性。结论 基于 PCF 的超连续谱光源可用于脑组织中钙动力学的同步成像和激发。泵浦 PCF 可以作为神经活动成像和激活的强大光源，并克服与多激光方法相关的有限光谱和空间。