We report a technologically novel microscopy system for bioimaging based on a 100 fs titanium:sapphire (Ti:Sa) laser pumped coherent continuum from a tailored, 9-cm long, all normal dispersion (ANDi) fiber, enabling concurrent image contrast with (a) spectral focusing coherent anti-Stokes Raman scattering (SF-CARS) (spanning 900–3200 cm −1 ) and (b) sum frequency generation (SFG). Both modalities were efficiently excited with power levels at the microscope focus compatible with biological samples. Moreover, using the continuum, images were recorded in the back-scattering (epi-detection) geometry, without the necessity for an expensive, computer-controlled, spatial light modulator (SLM), clearly demonstrating the strong signal levels achieved. Image contrast from the multiple modalities provided greater chemical and structural insights than imaging with any single technique in isolation. Numerical simulations supported these developments in regard to both the optimum fiber length for SC generation and the achievement of high spectral resolution in SF-CARS via careful group delay dispersion matching across the pump and Stokes pulses using just an inexpensive sequence of short glass blocks inserted into the Stokes beam. We show bio-images of mouse tissue recorded concurrently via label/stain-free contrast from multiple modalities: CARS, two-photon auto-fluorescence (TPaF) and second harmonic/sum frequency generation (SHG/SFG). Overall, our approach delivers optimum performance in back-scattered (epi-) detection configuration, suited for thick samples, at reduced complexity and cost. The addition of this simple fiber add-on to lasers already widely used for TPF microscopy can thus extend the capabilities of a significant number of existing microscopy laboratories.

中文翻译：

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多模态光谱聚焦 CARS 和 SFG 显微镜，具有来自微结构光纤的定制相干连续谱

我们报告了一种技术新颖的生物成像显微镜系统，该系统基于 100 fs 钛：蓝宝石 (Ti:Sa) 激光泵浦相干连续谱，来自定制的 9 厘米长全法向色散 (ANDi) 光纤，可实现与（a ) 光谱聚焦相干反斯托克斯拉曼散射 (SF-CARS)（跨越 900–3200 cm -1 ）和 (b) 和频生成 (SFG)。在与生物样品兼容的显微镜焦点处，两种模式都被有效激发。此外，使用连续介质，图像以背向散射（外延检测）几何结构记录，无需昂贵的计算机控制空间光调制器 (SLM)，清楚地展示了所实现的强信号水平。与单独使用任何单一技术的成像相比，来自多种模式的图像对比度提供了更多的化学和结构洞察力。数值模拟支持这些发展，即用于 SC 生成的最佳光纤长度和通过泵浦和斯托克斯脉冲之间的仔细群延迟色散匹配实现 SF-CARS 中的高光谱分辨率，仅使用插入的廉价短玻璃块序列斯托克斯梁。我们展示了通过来自多种模式的标记/无染色对比同时记录的小鼠组织的生物图像：CARS、双光子自发荧光 (TPaF) 和二次谐波/和频生成 (SHG/SFG)。总体而言，我们的方法在反向散射 (epi-) 检测配置中提供了最佳性能，适用于厚样品，同时降低了复杂性和成本。