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Single-Molecule Fluorescence Defines a New Dye Chemistry

Year: 2025

Type: article

Abstract: Century-old fluorescent dyes have regained life in the wave of super-resolution microscopy. The achievement of single-molecule resolution transcends conventional ensemble averaging, imposing specific photophysical requirements on fluorescent dyes at a millisecond temporal resolution and ∼1 nm single-molecule spatial precision. This review aims to synthesize the fundamental principle governing single-molecule fluorescence in terms of single-molecule photon flux, switching kinetics, and the structure-function correlations. It emphasizes the role of structural modulation down to the atomic level for tuning single-molecule fluorescence characteristics to enhance imaging resolutions and proposes a taxonomy of landmark structural modifications for regulating single-molecule fluorescence. The analysis would provide guidelines for ongoing dye engineering efforts to develop next-generation dyes.

Source: Chemical & Biomedical Imaging

DOI: 10.1021/cbmi.5c00148

Authors: Lujia Yang, Ying Zheng, Zhiwei Ye, Yi Xiao

Institutions: Dalian University of Technology, State Key Laboratory of Fine Chemicals, Dalian Medical University

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: diamond

Funders: National Natural Science Foundation of China, Department of Science and Technology of Liaoning Province, Dalian Medical University