Hu Guorong, Greene Joseph, Zhu Jiabei, Yang Qianwan, Zheng Shuqi, Li Yunzhe, Alido Jeffrey, Guo Ruipeng, Mertz Jerome, Tian Lei
Department of Electrical & Computer Engineering, Boston University , Boston, Massachusetts 02215, USA.
Currently with the Department of Electrical Engineering & Computer Sciences, University of California, Berkeley, California 94720, USA.
Biomed Opt Express. 2024 Jun 4;15(7):4101-4110. doi: 10.1364/BOE.527264. eCollection 2024 Jul 1.
HiLo microscopy is an optical sectioning structured illumination microscopy technique based on computationally combining two images: one with uniform illumination and the other with structured illumination. The most widely used structured illumination in HiLo microscopy is random speckle patterns, due to their simplicity and resilience to tissue scattering. Here, we present a novel HiLo microscopy strategy based on random patterns. Building on an off-the-shelf diffuser and a low-coherence LED source, we demonstrate that caustic HiLo can achieve 4.5 µm optical sectioning capability with a 20× 0.75 NA objective. In addition, with the distinct intensity statistical properties of caustic patterns, we show that our caustic HiLo outperforms speckle HiLo, achieving enhanced optical sectioning capability and preservation of fine features by imaging scattering fixed brain sections of 100 µm, 300 µm, and 500 µm thicknesses. We anticipate that this new structured illumination technique may find various biomedical imaging applications.
高-低显微镜是一种光学切片结构照明显微镜技术，它通过计算组合两张图像来实现：一张是均匀照明的图像，另一张是结构照明的图像。在高-低显微镜中，由于其简单性和对组织散射的耐受性，最广泛使用的结构照明是随机散斑图案。在这里，我们提出了一种基于随机图案的新型高-低显微镜策略。基于现成的漫射器和低相干发光二极管光源，我们证明了焦散高-低显微镜使用20×0.75 NA物镜时可实现4.5 µm的光学切片能力。此外，由于焦散图案具有独特的强度统计特性，我们表明我们的焦散高-低显微镜优于散斑高-低显微镜，通过对厚度为100 µm、300 µm和500 µm的散射固定脑切片成像，实现了增强的光学切片能力并保留了精细特征。我们预计这种新的结构照明技术可能会在各种生物医学成像应用中找到用武之地。
