Herein, we reported a simple, fast, and quantitative theoretical descriptor ΔGC-O that allows accurate predictions of a wide range of spontaneously blinking rhodamines. ΔGC-O denotes the Gibbs free energy differences between the closed and open forms of rhodamines and has a good linear relationship with experimental pKcycl values. This correlation affords an effective guide for the quantitative designs of spontaneously blinking rhodamines and eliminates trial-and-error. We have validated the predictive power of ΔGC-O via the development of two spontaneously blinking rhodamines of different colors and enhanced brightness. We also demonstrated their super-resolution imaging utilities in dynamic live-cell imaging. We expect that ΔGC-O will greatly facilitate the efficient creations of spontaneously blinking fluorophores and aid the advancements of super-resolution bioimaging techniques.
Keywords: Gibbs free energy; rhodamines; single-molecule localization microscopy; spontaneously blinking dyes; super-resolution imaging.
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