Yu Bin, Nie Mengjiao, Jiang Zizhen, Lin Danying, Qu Junle, Cao Huiqun
Opt Lett. 2025 Mar 15;50(6):2005-2008. doi: 10.1364/OL.555763.
Multifocal structured illumination microscopy (MSIM) provides a twofold resolution enhancement over the optical diffraction limit at depths of up to 50 μm in samples. This is achieved through sparse multifocal excitation patterns and digital image post-processing, making MSIM a highly advantageous technique for the three-dimensional super-resolution (SR) imaging of thick specimens. However, the spatial resolution of MSIM is inherently constrained by its underlying imaging principles. This paper presents what we believe to be a novel method that integrates SR optical fluctuation imaging based on Fourier ptychography and deconvolution (SFPD) with MSIM, termed SFPD-MSIM. Using photoblinking InP/ZnSe/ZnS core-shell quantum dot fluorescent probes for sample labeling, we demonstrate that, compared to wide-field imaging microscopy, SFPD-MSIM achieves fourfold resolution improvement. Additionally, it substantially reduces the image-acquisition time while preserving the structural integrity of the original samples. This advancement marks a major step forward in MSIM technology, providing a powerful tool for detailed structural analysis of complex and thick biological specimens.
多焦点结构照明显微镜（MSIM）在样品深度达50微米时，可将分辨率提高两倍，超越光学衍射极限。这是通过稀疏多焦点激发模式和数字图像后处理实现的，使MSIM成为厚样本三维超分辨率（SR）成像的极具优势的技术。然而，MSIM的空间分辨率本质上受其成像原理的限制。本文提出了一种我们认为新颖的方法，即将基于傅里叶叠层成像和反卷积的超分辨率光学涨落成像（SFPD）与MSIM相结合，称为SFPD-MSIM。使用光致闪烁的InP/ZnSe/ZnS核壳量子点荧光探针标记样品，我们证明，与宽场成像显微镜相比，SFPD-MSIM的分辨率提高了四倍。此外，它在保持原始样品结构完整性的同时，大幅缩短了图像采集时间。这一进展标志着MSIM技术向前迈出了重要一步，为复杂厚生物样本的详细结构分析提供了强大工具。
