论文标题
RSHG:重新扫描第二谐波生成显微镜
rSHG: Re-scan Second Harmonic Generation Microscopy
论文作者
论文摘要
第二次谐波生成显微镜(SHG)通常被公认为是无标签的3D可视化组织和高级材料的功能强大工具,其最受欢迎的应用之一是胶原蛋白成像。尽管巨大的需求,但在过去几年中,在基于荧光的光学纳米镜检查中,超级分辨SHG成像的进展滞后。在这项工作中,我们在胶原组织上进行了定量显示,通过将SHG成像与重新扫描显微镜的分辨率相结合,该分辨率超过了衍射极限,〜1.4倍可用。除了重新扫描的第二次谐波生成显微镜(RSHG)外,我们还证明了超级分辨的重新扫描两光激发荧光显微镜(RTPEF)。这两种技术是通过修改重新扫描共聚焦显微镜(RCM)来实现的,从而保留其初始功能,从而导致了多模式RSHG/RSHG/RTPEF/RCM系统。鉴于重新扫描显微镜的简单性和灵活性,我们认为所报告的结果可能会增加依赖超级解决非线性光学成像的应用的数量和性质。
Second Harmonic Generation Microscopy (SHG) is generally acknowledged as a powerful tool for the label-free 3D visualization of tissues and advanced materials, with one of its most popular applications being collagen imaging. Although the great need, progress in super-resolved SHG imaging lags behind the developments reported over the past years in fluorescence-based optical nanoscopy. In this work, we quantitatively show on collagenous tissues that by combining SHG imaging with re-scan microscopy resolutions that surpass the diffraction limit with ~1.4x become available. Besides Re-scan Second Harmonic Generation Microscopy (rSHG), we demonstrate as well super-resolved Re-scan Two-Photon Excited Fluorescence Microscopy (rTPEF). These two techniques are implemented by modifying a Re-scan Confocal Microscope (RCM), retaining its initial function, resulting thus in a multimodal rSHG/rTPEF/RCM system. Given the simplicity and flexibility of re-scan microscopy, we consider that the reported results are likely to augment the number and nature of applications relying on super-resolved non-linear optical imaging.