Advanced Fluorescence Microscopy Techniques Open access Peer reviewed

A series of spontaneously blinking dyes for super-resolution microscopy

Katie L. Holland, Sarah E. Plutkis, Timothy A. Daugird, Abhishek Sau and 15 more

Nature Methods | Apr 15, 2026 | 28 citations

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What this paper is about

Dyes with tuned on:off ratios that enable single-molecule localization microscopy and super-resolution optical fluctuation imaging of biomolecular structures in vitro and in cells are reported.

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Spontaneously blinking fluorophores toggle between nonfluorescent and fluorescent forms without caging groups or redox buffers, enabling super-resolution imaging. The intrinsic blinking of such dyes is governed by molecular structure and modulated by environment; there is no one-size-fits-all fluorophore suitable for every imaging context. We report dyes with tuned on:off ratios that enable single-molecule localization microscopy and super-resolution optical fluctuation imaging of biomolecular structures in vitro and in cells.

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Authors

Researchers on this paper

Katie L. Holland

first | Howard Hughes Medical Institute | ORCID 0009-0007-1183-195X

Sarah E. Plutkis

middle | Howard Hughes Medical Institute | ORCID 0009-0003-1822-0466

Timothy A. Daugird

middle | University of North Carolina at Chapel Hill | ORCID 0000-0002-6313-0246

Abhishek Sau

middle | University of North Carolina at Chapel Hill | ORCID 0000-0002-5560-5360

Jonathan B. Grimm

middle | Howard Hughes Medical Institute | ORCID 0000-0003-0331-4200

Brian P. English

middle | Howard Hughes Medical Institute | ORCID 0000-0002-4037-6294

Qinsi Zheng

middle | Howard Hughes Medical Institute

Sandeep Dave

middle | Texas A&M Health Science Center | ORCID 0000-0003-2832-2098

Fariha Rahman

middle | University of North Carolina at Chapel Hill | ORCID 0000-0001-8014-3025

Liangqi Xie

middle | Howard Hughes Medical Institute | ORCID 0000-0002-6546-9622

Peng Dong

middle | University of North Carolina at Chapel Hill | ORCID 0000-0003-4946-1409

Ariana N. Tkachuk

middle | Howard Hughes Medical Institute | ORCID 0000-0001-8991-9643

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Citation

BibTeX

@article{Holland2026series,
  title = {A series of spontaneously blinking dyes for super-resolution microscopy},
  author = {Katie L. Holland and Sarah E. Plutkis and Timothy A. Daugird and Abhishek Sau and Jonathan B. Grimm and Brian P. English and Qinsi Zheng and Sandeep Dave and Fariha Rahman and Liangqi Xie and Peng Dong and Ariana N. Tkachuk and Timothy A. Brown and Robert H. Singer and Zhe Liu and Catherine G. Galbraith and Siegfried M. Musser and Wesley R. Legant and Luke D. Lavis},
  journal = {Nature Methods},
  year = {2026},
  doi = {10.1038/s41592-026-03062-5},
  url = {https://doi.org/10.1038/s41592-026-03062-5}
}

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