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Deep-Tissue Two-Photon Imaging in Brain and Peripheral Nerve with a Compact High-Pulse Energy Ytterbium Fiber Laser

机译:紧凑型高脉冲能量Energy光纤激光器在脑和周围神经中的深组织双光子成像。

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Two-photon microscopy is a powerful tool of current scientific research, allowing optical visualization of structures below the surface of tissues. This is of particular value in neuroscience, where optically accessing regions within the brain is critical for the continued advancement in understanding of neural circuits. However, two-photon imaging at significant depths have typically used Ti:Sapphire based amplifiers that are prohibitively expensive and bulky. In this study, we demonstrate deep tissue two-photon imaging using a compact, inexpensive, turnkey operated Ytterbium fiber laser (Y-Fi, KM Labs). The laser is based on all-normal dispersion (ANDi) that provides short pulse durations and high pulse energies. Depth measurements obtained in ex vivo mouse cortex exceed those obtainable with standard two-photon microscopes using Ti:Sapphire lasers. In addition to demonstrating the capability of deep-tissue imaging in the brain, we investigated imaging depth in highly-scattering white matter with measurements in sciatic nerve showing limited optical penetration of heavily myelinated nerve tissue relative to grey matter.
机译:双光子显微镜是当前科学研究的强大工具,可对组织表面以下的结构进行光学可视化。这在神经科学中具有特别的价值,在大脑中,光学进入大脑区域对于持续理解神经回路至关重要。然而,在相当大的深度处的双光子成像通常已经使用了基于Ti:Sapphire的放大器,价格昂贵且体积庞大。在这项研究中,我们展示了使用紧凑,廉价,交钥匙操作的tter光纤激光器(Y-Fi,KM Labs)进行的深层组织双光子成像。激光器基于全常态色散(ANDi),可提供短脉冲持续时间和高脉冲能量。在离体小鼠皮层中获得的深度测量值超过了使用Ti:Sapphire激光器的标准双光子显微镜可获得的测量值。除了证明大脑中深层组织成像的能力外,我们还通过在坐骨神经中进行测量,调查了高度散射白质的成像深度,结果显示相对于灰质,重度有髓神经组织的光学穿透力有限。

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