Confocal Raman microscopy, a highly specific and label-free technique for the microscale study of thick samples, often presents difficulties due to weak Raman signals. Inhomogeneous samples introduce wavefront aberrations that further reduce these signals, requiring even longer acquisition times. In this study, we introduce Adaptive Optics to confocal Raman microscopy for the first time to counteract such aberrations, significantly increasing the Raman signal and image quality. The method is designed to integrate seamlessly with existing commercial microscopes without hardware modifications. It uses a wavefront sensorless approach to measure aberrations using an optofluidic, transmissive spatial light modulator that can be attached to the microscope nosepiece. Our experimental results demonstrate the compensation of aberrations caused by artificial scatterers and mouse brain tissue, improving spatial resolution and achieving up to 3.5-fold signal enhancements. Our results provide a basis for the molecular label-free study of biological systems at greater imaging depths.

中文翻译：

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具有自适应光学元件的共聚焦拉曼显微镜


共聚焦拉曼显微镜是一种高度特异性且无标记的技术，用于厚样品的微量研究，由于拉曼信号较弱，经常存在困难。不均匀的样品会引入波前像差，从而进一步降低这些信号，因此需要更长的采集时间。在这项研究中，我们首次将自适应光学技术引入共聚焦拉曼显微镜，以抵消此类像差，显著提高拉曼信号和图像质量。该方法旨在与现有的商用显微镜无缝集成，无需修改硬件。它使用无波前传感器方法，使用可连接到显微镜物镜转换器的光流体透射空间光调制器来测量像差。我们的实验结果表明，可以补偿人工散射体和小鼠脑组织引起的像差，提高空间分辨率并实现高达 3.5 倍的信号增强。我们的结果为在更大成像深度下对生物系统进行分子无标记研究提供了基础。