Time-resolved photon counting Fourier-transform micro-spectroscopy enables simultaneous Raman and fluorescence lifetime imaging

Shang, Lindong, Bao, Xiaodong, Peng, Hao, Chen, Fuyuan, Wang, Yu, Liu, Kunxiang, Liang, Peng, Wang, Yuntong, Tang, Xusheng, Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023 and Li, Bei 2025. Time-resolved photon counting Fourier-transform micro-spectroscopy enables simultaneous Raman and fluorescence lifetime imaging. Light: Science & Applications 14 (1) , 378. 10.1038/s41377-025-02020-8

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Abstract

To address the current limitations of time-gated Raman spectroscopy, specifically its narrow spectral range and low spectral resolution, and simultaneously acquire Raman and fluorescence life-time images, we have developed a Fourier-transform photon counting spectroscopy platform. A Mach-Zehnder interferometer employing a high accuracy linear motor stage was combined with photon-counting avalanche diodes and time-tagged acquisition, allowing to sort photons into a matrix of stage positions determined using their coarse arrival time with 50 ns steps of the excitation laser repetition period, and a fine arrival time of 80 ps resolution relative to the excitation pulse of 100 ps duration. The instrument achieves a time resolution of 547 ps, a wide spectral range of −1000 to 10,000 cm−1 Raman shift from the excitation at 532 nm wavelength, and a high spectral resolution of 0.05 cm−1. For experimental validation, we used fluorescently coated silicon wafers and fluorescent plastic microspheres. Raman signal was observed during the laser excitation pulse within the time-resolution, while fluorescence signals dominate afterwards. The results confirm that the instrument can effectively separate Raman and fluorescence signals.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Schools > Physics and Astronomy Schools > Biosciences
Additional Information:	License information from Publisher: LICENSE 1: URL: http://creativecommons.org/licenses/by/4.0/, Type: open-access
Publisher:	Springer Nature [academic journals on nature.com]
Date of First Compliant Deposit:	4 November 2025
Date of Acceptance:	19 August 2025
Last Modified:	04 Nov 2025 11:31
URI:	https://orca.cardiff.ac.uk/id/eprint/182098

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