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Updated: Oct 10, 2025

Author Spotlight: Integrated OPTIR-FISH for Single-Cell Metabolic and Identity Analysis in Complex Environments
Published on: February 23, 2024
Halftone spatial frequency domain imaging enables kilohertz high-speed label-free non-contact quantitative mapping of
Yanyu Zhao1, Bowen Song2, Ming Wang3
1Beijing Advanced Innovation Center for Biomedical Engineering, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Engineering Medicine, and with the School of Biological Science and Medical Engineering, Beihang University, 100191, Beijing, China. yanyuzhao@buaa.edu.cn.
Abstract:
The ability to quantify optical properties (i.e., absorption and scattering) of strongly turbid media has major implications on the characterization of biological tissues, fluid fields, and many others. However, there are few methods that can provide wide-field quantification of optical properties, and none is able to perform quantitative optical property imaging with high-speed (e.g., kilohertz) capabilities. Here we develop a new imaging modality termed halftone spatial frequency domain imaging (halftone-SFDI), which is approximately two orders of magnitude faster than the state-of-the-art, and provides kilohertz high-speed, label-free, non-contact, wide-field quantification for the optical properties of strongly turbid media. This method utilizes halftone binary patterned illumination to target the spatial frequency response of turbid media, which is then mapped to optical properties using model-based analysis. We validate the halftone-SFDI on an array of phantoms with a wide range of optical properties as well as in vivo human tissue. We demonstrate with an in vivo rat brain cortex imaging study, and show that halftone-SFDI can longitudinally monitor the absolute concentration as well as spatial distribution of functional chromophores in tissue. We also show that halftone-SFDI can spatially map dual-wavelength optical properties of a highly dynamic flow field at kilohertz speed. Together, these results highlight the potential of halftone-SFDI to enable new capabilities in fundamental research and translational studies including brain science and fluid dynamics.

