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Spontaneously Blinking Rhodamine Dyes for Single-Molecule Localization Microscopy.

Weijie Chi1,2, Davin Tan2, Qinglong Qiao3

  • 1Collaborative Innovation Center of One Health, School of Science, Hainan University, Renmin Road 58, Haikou, 570228, P. R. China.

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PubMed
Summary

This review details spontaneously blinking rhodamine fluorophores, crucial for advanced single-molecule localization microscopy (SMLM). It covers their development and mechanisms, aiding super-resolution imaging progress.

Keywords:
Rhodamine DyesSingle-Molecule Localization MicroscopySpirocyclization EquilibriumSpontaneously Blinking Fluorophores

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Area of Science:

  • Chemistry and Biology
  • Microscopy and Imaging Technologies

Background:

  • Single-molecule localization microscopy (SMLM) is vital for biological and chemical imaging.
  • Fluorophores are essential for SMLM, enabling super-resolution fluorescence imaging.
  • Spontaneously blinking fluorophores simplify SMLM experiments and extend imaging times.

Purpose of the Study:

  • To provide a comprehensive review of spontaneously blinking rhodamine development from 2014-2023.
  • To elucidate the key mechanistic aspects of intramolecular spirocyclization reactions in these fluorophores.
  • To offer design guidelines for advancing super-resolution imaging technologies.

Main Methods:

  • Literature review of scientific publications from 2014 to 2023.
  • Analysis of mechanistic studies on intramolecular spirocyclization reactions.
  • Synthesis of design principles for spontaneously blinking fluorophores.

Main Results:

  • Overview of the evolution of spontaneously blinking rhodamines over the past decade.
  • Detailed explanation of the photophysical mechanisms, particularly intramolecular spirocyclization.
  • Identification of key design strategies for improved fluorophore performance.

Conclusions:

  • Spontaneously blinking rhodamines have significantly advanced SMLM capabilities.
  • Understanding spirocyclization mechanisms is key to designing next-generation fluorophores.
  • This review provides a foundation for future innovations in super-resolution microscopy.