Excitation spectral microscopy for highly multiplexed fluorescence imaging and quantitative biosensing

Kun Chen; Rui Yan; Limin Xiang; Ke Xu

doi:10.1117/12.2612265

2 March 2022 Excitation spectral microscopy for highly multiplexed fluorescence imaging and quantitative biosensing

Kun Chen, Rui Yan, Limin Xiang, Ke Xu

Author Affiliations +

Proceedings Volume PC11971, High-Speed Biomedical Imaging and Spectroscopy VII; PC119710A (2022) https://doi.org/10.1117/12.2612265
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

We discuss our recent efforts to achieve highly multiplexed fast fluorescence imaging and quantitative biosensing through camera frame-synchronized scanning of the excitation wavelength in the wide field. For live-cell microscopy, we thus attain low (~1%) crosstalks and ~10 ms temporal resolutions for up to six fluorophores via linear unmixing, and further develop novel, quantitative imaging schemes for both bi-state and FRET fluorescent biosensors. These capabilities are further integrated to multiplex absolute pH imaging with three additional target proteins in the mitophagy pathway. Together, excitation spectral microscopy provides exceptional opportunities for highly multiplexed fluorescence imaging without fluorescence dispersion.

Conference Presentation

Citation Download Citation

Kun Chen, Rui Yan, Limin Xiang, and Ke Xu "Excitation spectral microscopy for highly multiplexed fluorescence imaging and quantitative biosensing", Proc. SPIE PC11971, High-Speed Biomedical Imaging and Spectroscopy VII, PC119710A (2 March 2022); https://doi.org/10.1117/12.2612265

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