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Annual Review of Physical Chemistry

Volume 76, 2025

Merging Vibrational Spectroscopy with Fluorescence Microscopy: Combining the Best of Two Worlds

  • Naixin Qian1, Hanqing Xiong2, Lu Wei3, Lixue Shi4 and Wei Min1
  • 1Department of Chemistry, Columbia University, New York, NY, USA; email: [email protected], [email protected] 2National Biomedical Imaging Center, College of Future Technology, Peking University, Beijing, China; email: [email protected] 3Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, USA; email: [email protected] 4Shanghai Xuhui Central Hospital, Zhongshan-Xuhui Hospital, and Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism, Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, Shanghai, China; email: [email protected]
  • Vol. 76:279-301 (Volume publication date April 2025)
  • First published as a Review in Advance on February 03, 2025
  • Copyright © 2025 by the author(s).
    This work is licensed under a Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. See credit lines of images or other third-party material in this article for license information.

Abstract

Vibrational spectroscopy and fluorescence spectroscopy have historically been two established but separate fields of molecular spectroscopy. While vibrational spectroscopy provides exquisite chemical information, fluorescence spectroscopy often offers orders of magnitude higher detection sensitivity. However, they each lack the advantages of each other. In recent years, a series of novel nonlinear optical spectroscopy studies have been developed that merge both spectroscopies into a single double-resonance process. These techniques combine the chemical specificity of Raman or infrared (IR) spectroscopy with the superb detection sensitivity and spatial resolution of fluorescence microscopy. Many facets have been explored, including Raman transition versus IR transition, time domain versus frequency domain, and spectroscopy versus microscopy. Notably, single-molecule vibrational spectroscopy has been achieved at room temperature without the need for plasmonics. Even superresolution vibrational imaging beyond the diffraction limit was demonstrated. This review summarizes the growing field of vibrational-encoded fluorescence microscopy, including key technical developments, emerging applications, and future prospects.

Keyword(s): double-resonance spectroscopy, fluorescence microscopy, nonlinear spectroscopy, optical imaging, single-molecule spectroscopy, vibrational spectroscopy, vibrational-encoded fluorescence microscopy
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2025-04-21
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/content/journals/10.1146/annurev-physchem-082423-121033
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Merging Vibrational Spectroscopy with Fluorescence Microscopy: Combining the Best of Two Worlds
Annual Review of Physical Chemistry 76, 279 (2025); https://doi.org/10.1146/annurev-physchem-082423-121033
/content/journals/10.1146/annurev-physchem-082423-121033
/content/journals/10.1146/annurev-physchem-082423-121033
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