Zhang Huaiyuan, Hu Yiting, Pu Xingwei, Zhang Shizheng, He Yi, Chen Kun, Liu Ziji
University of Electronic Science and Technology of China, School of Optoelectronic Science and Engineering, Chengdu, China.
J Biomed Opt. 2025 Apr;30(4):046501. doi: 10.1117/1.JBO.30.4.046501. Epub 2025 Apr 8.
The multiplexing capabilities of fluorescence imaging are enhanced by its exceptional molecular specificity with diverse fluorescent probes, making it a powerful tool for studying complex biological structures, organization, and functions. Recent advances in super-resolution fluorescence microscopy have further revolutionized our ability to explore biology and related fields. However, current multicolor super-resolution fluorescence imaging systems often come with high costs and bulky designs.
We present a multicolor extended resolution fluorescence imaging system that uses light-emitting diode to simplify the optical path, make the design more compact, and reduce system costs.
This multicolor extended resolution fluorescence imaging system is based on structured illumination, utilizing a simple diffraction unit positioned between the light source and the sample in a wide-field microscope. Notably, this design could be easily integrated into standard widefield microscopes as a convenient add-on unit, enabling extended resolution imaging.
Our system demonstrates concurrent extended resolved imaging of three-color microsphere beads and successfully showcases multicolor extended resolution fluorescence imaging of biological tissue samples, revealing intricate structural details.
This system provides a structurally simple, cost-effective alternative to traditional microscopes, offering flexible multicolor extended resolution fluorescence imaging and potential applications in multimodal imaging.
荧光成像的复用能力因其使用多种荧光探针而具有出色的分子特异性而得到增强，使其成为研究复杂生物结构、组织和功能的强大工具。超分辨率荧光显微镜的最新进展进一步彻底改变了我们探索生物学及相关领域的能力。然而，当前的多色超分辨率荧光成像系统往往成本高昂且设计笨重。
我们提出一种多色扩展分辨率荧光成像系统，该系统使用发光二极管来简化光路，使设计更紧凑，并降低系统成本。
这种多色扩展分辨率荧光成像系统基于结构照明，在宽视场显微镜中利用位于光源和样品之间的一个简单衍射单元。值得注意的是，这种设计可以很容易地作为一个方便的附加单元集成到标准宽视场显微镜中，实现扩展分辨率成像。
我们的系统展示了三色微球珠的并发扩展分辨率成像，并成功展示了生物组织样品的多色扩展分辨率荧光成像，揭示了复杂的结构细节。
该系统为传统显微镜提供了一种结构简单、成本效益高的替代方案，提供了灵活的多色扩展分辨率荧光成像以及在多模态成像中的潜在应用。
