A series of spontaneously blinking dyes for super-resolution microscopy

Katie L. Holland; Sarah E. Plutkis; Brian P. English; Timothy A. Daugird; Abhishek Sau; Jonathan B. Grimm; Qinsi Zheng; Ankith Sharma; Sandeep Dave; Anja Schmidt; Fariha Rahman; Liangqi Xie; Peng Dong; Ariana N. Tkachuk; Timothy A. Brown; Robert H. Singer; Zhe J. Liu; Siegfried M. Musser; Wesley R. Legant; Catherine G. Galbraith; Luke D. Lavis

doi:10.1038/s41592-026-03062-5

Cell BiologyNature MethodsApr 15, 2026

Peer-reviewed journal article

A series of spontaneously blinking dyes for super-resolution microscopy

Katie L. Holland iD, Sarah E. Plutkis iD, Brian P. English iD, Timothy A. Daugird iD, Abhishek Sau iD, Jonathan B. Grimm iD, Qinsi Zheng, Ankith Sharma iD, Sandeep Dave iD, Anja Schmidt iD, Fariha Rahman iD, Liangqi Xie iD, Peng Dong iD, Ariana N. Tkachuk iD, Timothy A. Brown iD, Robert H. Singer iD, Zhe J. Liu iD, Siegfried M. Musser iD, Wesley R. Legant iD, Catherine G. Galbraith iD, Luke D. Lavis iD

Abstract

Spontaneously blinking fluorophores permit the detection and localization of individual molecules without reducing buffers or caging groups, thus simplifying single-molecule localization microscopy (SMLM). The intrinsic blinking properties of such dyes are dictated by molecular structure and modulated by environment, which can limit utility. We report a series of tuned spontaneously blinking dyes with duty cycles that span two orders of magnitude, allowing facile SMLM in cells and dense biomolecular structures.

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