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Dryad

Electrochemical fluorescence modulation enables simultaneous multicolor imaging

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Mar 19, 2025 version files 45.16 MB

Abstract

We present a new multicolor imaging strategy on a standard fluorescence microscope, where up to four fluorophores with high spectra overlap can be resolved with a single-color optical configuration. Under electrochemical modulation, the fluorophores are regulated between bright states and dim states, with each displaying a distinct fluorescence response pattern. These unique fluorescence-potential profiles enable effective separation of different fluorophores through linear unmixing. We also demonstrated that electrochemical fluorescence switching is readily applicable for multicolor STED imaging. With no modification to the optical setups and easy adaptation to different microscopes, we anticipate that color unmixing based on electrochemical fluorescence switching will provide an easily accessible multicolor imaging pathway for discoveries in diverse fields.