Implementing caustic illumination in HiLo microscopy

Guorong Hu; Joseph Greene; Jiabei Zhu; Qianwan Yang; Yunzhe Li; Jeffrey Alido; Ruipeng Guo; Lei Tian

doi:10.1117/12.2692805

13 March 2024 Implementing caustic illumination in HiLo microscopy

Guorong Hu, Joseph Greene, Jiabei Zhu, Qianwan Yang, Yunzhe Li, Jeffrey Alido, Ruipeng Guo, Lei Tian

Proceedings Volume PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI; PC128480J (2024) https://doi.org/10.1117/12.2692805
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

HiLo microscopy is a widefield optical sectioning technique that involves computational reconstruction from two images, one with structured illumination and the other with uniform illumination. A variety of methods, including speckle and periodic grids, can be employed to achieve structured illumination. In this study, we introduce a novel HiLo strategy that utilizes an off-the-shelf holographic diffuser and a low-coherence LED source to generate random caustic patterns. This method offers several benefits over existing ones, such as simplicity and cost-effectiveness. We achieve 4.5 µm optical sectioning capability with a 20x 0.75 NA objective and demonstrate the performance of our method by imaging a 400 µm thick, highly scattering brain section. We anticipate that our caustic-based structured illumination approach will augment the versatility of HiLo microscopy and extend to various imaging applications.

Conference Presentation

Citation Download Citation

Guorong Hu, Joseph Greene, Jiabei Zhu, Qianwan Yang, Yunzhe Li, Jeffrey Alido, Ruipeng Guo, and Lei Tian "Implementing caustic illumination in HiLo microscopy", Proc. SPIE PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, PC128480J (13 March 2024); https://doi.org/10.1117/12.2692805

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KEYWORDS

Light sources and illumination

Microscopy

Cesium

Linear filtering

Optical transfer functions

Diffusers

Fluorescence

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