Early View e202500187
RESEARCH ARTICLE

Multiparametric Wide-Field Fluorescence Imaging via Polarization Modulation With Liquid Crystal Rotators

Wenshuai Li

Wenshuai Li

School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, China

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Bin Xu

Bin Xu

School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, China

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Chaofu Sun

Chaofu Sun

School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, China

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Weiping Liu

Weiping Liu

Xi'an Institute of Applied Optics, Xi'an, China

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Yang Zhang

Yang Zhang

School of Artificial Intelligence, Luoyang Institute of Science and Technology, Luoyang, China

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Ji Wu

Ji Wu

Renji Hospital, Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Xuefeng Liu

Xuefeng Liu

School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, China

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Jichuan Xiong

Corresponding Author

Jichuan Xiong

School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, China

Correspondence:

Jichuan Xiong ([email protected])

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First published: 15 July 2025

Funding: This work was supported by the National Major Scientific Instruments and Equipment Development Project under Grant No. 61827814 and the Ministry of Education Collaborative Project (B17023).

Wenshuai Li and Bin Xu contributed equally to the manuscript.

ABSTRACT

Fluorescence polarization imaging provides critical insights into molecular orientation, yet existing methods face limitations in parameter extraction efficiency and implementation complexity. This study proposes Wide-Field Multiparametric Fluorescence Imaging (WMPFI) using a Liquid Crystal Polarization Rotator (LCPR) for rapid polarization state modulation that generates pixel-level intensity modulations that encode fluorophore orientation. By analyzing fluorescence intensity variations under different polarization excitations, WMPFI reconstructs sample structural information through parametric imaging without requiring optical lock-in detection or computational reconstruction algorithms. Comparative experiments with Conventional Microscopy (CM) demonstrate WMPFI's enhanced sensitivity to anisotropic fluorescent dipole orientations, achieving superior contrast and resolution in imaging neural stem cells and skin tissues. The method's capacity for multi-parameter acquisition through polarization modulation offers a simplified approach for probing subcellular material exchange dynamics, with potential extensions to super-resolution imaging modalities.

Conflicts of Interest

The authors declare no conflicts of interest.

Data Availability Statement

The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions.

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