Descriptor ΔGC‐O Enables the Quantitative Design of Spontaneously Blinking Rhodamines for Live‐Cell Super‐Resolution Imaging

Abstract Herein, we reported a simple, fast, and quantitative theoretical descriptor Δ G C‐O that allows accurate predictions of a wide range of spontaneously blinking rhodamines. Δ G C‐O denotes the Gibbs free energy differences between the closed and open forms of rhodamines and has a good linear relationship with experimental p K cycl 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 Δ G C‐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 Δ G C‐O will greatly facilitate the efficient creations of spontaneously blinking fluorophores and aid the advancements of super‐resolution bioimaging techniques.