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We introduce a strategy to optimize the photoswitching behavior of rhodamines for (d)STORM super‐resolution microscopy. By replacing the benzene ring in the rhodamine core with a permanently charged 1,3‐disubstituted imidazolium, the resultant dyes are markedly sensitized toward photoswitching, and exhibit outstanding (d)STORM performance with fast on‐off switching, long‐lasting blinking, and bright single‐molecule emission. We thus attain excellent (d)STORM images under green excitation that are on par with the “ideal” red‐excited dyes, including for difficult structures such as the mammalian actin cytoskeleton, and demonstrate high‐quality two‐color three‐dimensional (d)STORM.

We introduce a strategy to optimize the photoswitching behavior of rhodamines for (d)STORM super‐resolution microscopy. By replacing the benzene ring in the rhodamine core with a permanently charged 1,3‐disubstituted imidazolium, the resultant dyes are markedly sensitized toward photoswitching, and exhibit outstanding (d)STORM performance with fast on‐off switching, long‐lasting blinking, and bright single‐molecule emission. We thus attain excellent (d)STORM images under green excitation that are on par with the “ideal” red‐excited dyes, including for difficult structures such as the mammalian actin cytoskeleton, and demonstrate high‐quality two‐color three‐dimensional (d)STORM.

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