Single-molecule localization microscopy, which is one of the most widely used super-resolution microscopy techniques, relies on the photoswitching of fluorophores. According to the photoswitching mechanism proposed by Doory Kim et al. in their Research Article (e202405246), the rate of switch-off is determined by the nucleophilicity of the photoswitching reagent, whereas the rate of switch-on, which is a two-photon-induced dissociation process, is determined by the bond dissociation energy of the photoswitching reagent–fluorophore pair.
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Go et al. (2024) studied this question.
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