AI 中文总结
本文提出ISES纳米镜技术，在200μm深度实现37nm分辨率，平衡了深层组织成像的分辨率、深度与光毒性，兼容多种饱和探针，为深层组织超分辨成像及纳米探针开发提供了新方案。
AI 中文摘要
深层组织超分辨成像的发展是实现无创活体光学观测的重要桥梁，但在空间分辨率、成像深度与光毒性的平衡方面仍存在挑战。本文提出一种名为图像扫描发射饱和（Image Scanning Emission Saturation, ISES）的纳米镜技术，在200μm成像深度下实现了37纳米的横向分辨率，相当于激发波长的1/25。该技术采用976纳米环形激发光束，结合成像扫描显微镜配置，应用基于饱和效应的点扩散函数（Point Spread Function, PSF）工程和像素级共聚焦针孔增强来提升空间分辨率。由于图像光学传递函数（Optical Transfer Function, OTF）的高频与低频分量可在单次扫描中通过不同相机像素同步采集，因此可利用单个扫描数据集进行傅里叶域融合以进一步提升图像质量。与传统基于环形激发光束的自适应像素重分配方法相比，本策略保留了原始频率分布，减少了复杂生物样品成像中的重建伪影。该策略具有通用性，可兼容多种表现出饱和行为的探针，不仅为深层组织超分辨成像提供了一种通用且实用的方法，还为下一代成像纳米探针的开发提供了思路。
英文摘要
The development of deep-tissue super-resolution imaging serves as an essential bridge toward non-invasive in vivo optical observation. However, there remain challenges to balance spatial resolution, imaging depth and phototoxicity. Here, we present a nanoscopy namely Image Scanning Emission Saturation (ISES) nanoscopy, achieving a lateral resolution of 37 nm, 1/25th of the excitation wavelength, at an imaging depth of 200 μm. Using a 976-nm doughnut-shaped excitation beam within an imaging-scanning microscopy configuration, we apply saturation-based point spread function (PSF) engineering and pixel-level confocal-pinhole enhancement to improve spatial resolution. As the high- and low-frequency components of the image OTF are concurrently acquired in a single scan via different camera pixels, Fourier-domain fusion can be employed with a single scanning dataset to further improve image quality. Compared with the traditional doughnut excitation beam-based adaptive pixel reassignment method, our strategy preserves the original frequency distributions and mitigates reconstruction artifacts in complex biological sample imaging. This strategy is generalizable and compatible with a variety of probes displaying saturation behavior. Beyond enabling a versatile and practical approach for deep tissue super-resolution imaging, it also informs the development of next-generation nanoprobes for imaging.
Comments16 pages,5 figures
