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Biocompatible TADF Probes for Highly Multiplexed Fluorescence Lifetime Imaging.

Pilar Suárez de Cepeda1,2,3, Ferran Nadal-Bufi1,2, Janine Haug4,5,6

  • 1Centre for Inflammation Research, The University of Edinburgh, Edinburgh, EH16 4UU, UK.

Angewandte Chemie (International Ed. in English)
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Summary
This summary is machine-generated.

New thermally activated delayed fluorescence (TADF) nanoprobes offer long, diverse fluorescence lifetimes for multiplexed imaging. This enables simultaneous imaging of five probes in a single spectral window using fluorescence lifetime imaging microscopy (FLIM).

Keywords:
BacteriaFluorescence lifetime imaging microscopy (FLIM)FluorophoresNanoprobes

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Area of Science:

  • Biophysics
  • Optical Imaging
  • Materials Science

Background:

  • Fluorescence lifetime imaging microscopy (FLIM) offers sensitive cellular microenvironment analysis.
  • Conventional fluorophores have limited lifetimes (<5 ns), restricting multiplexed imaging capabilities.
  • Longer, diverse fluorescence lifetimes are needed for advanced FLIM applications.

Purpose of the Study:

  • To design and evaluate novel thermally activated delayed fluorescence (TADF) nanoprobes for multiplexed FLIM.
  • To develop nanoprobes with extended and varied fluorescence lifetimes suitable for biological imaging.
  • To demonstrate the potential of TADF emitters for advanced multi-color FLIM.

Main Methods:

  • Systematic design and synthesis of 36 biocompatible TADF nanoprobes.
  • Evaluation of nanoprobes in aqueous media for fluorescence lifetime characteristics (up to 15 ns).
  • Application of selected TADF probes for live-cell imaging of bacterial cells under physiological conditions.
  • Utilized FLIM-phasor analysis for simultaneous multi-probe imaging within a single spectral window.

Main Results:

  • Successfully synthesized 36 TADF nanoprobes with long and diverse fluorescence lifetimes.
  • Demonstrated live-cell imaging of bacterial cells using these novel nanoprobes.
  • Achieved unprecedented simultaneous imaging of five nanoprobes in one spectral window via FLIM-phasor strategy.
  • Validated the broad range of fluorescence lifetimes of TADF emitters for multiplexed imaging.

Conclusions:

  • Thermally activated delayed fluorescence (TADF) emitters are highly effective scaffolds for developing advanced FLIM probes.
  • The developed TADF nanoprobes significantly expand the possibilities for multiplexed imaging in biological studies.
  • This work unlocks the potential of FLIM for multi-color biological investigations by overcoming limitations of conventional fluorophores.