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Total Internal Reflection Fluorescence Microscopy01:05

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Multi-Level Confinement Single-Molecule Charge Transfer Activated PRET for Near-Infrared Targeted Cell Imaging.

Zhuo Lei1, Sai Li1, Pei-Ao Sun1

  • 1College of Chemistry, State Key Laboratory of Elemento-Organic Chemistry, Nankai University, Tianjin, P. R. China.

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Summary
This summary is machine-generated.

This study introduces a novel phosphorescent resonance energy transfer (PRET) system using supramolecular assembly for advanced bioimaging. The system achieves near-infrared delayed fluorescence, overcoming traditional limitations for enhanced cellular visualization.

Keywords:
cell imagingcharge transfermulti‐level confinementnear‐infrared delayed fluorescence emissionphosphorescent resonance energy transfer

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Area of Science:

  • Supramolecular Chemistry
  • Bioimaging
  • Materials Science

Background:

  • Phosphorescent resonance energy transfer (PRET) typically has short-wavelength limitations.
  • Developing efficient PRET systems for near-infrared (NIR) bioimaging remains a challenge.
  • Supramolecular assemblies offer tunable properties for advanced optical applications.

Purpose of the Study:

  • To design and characterize a novel supramolecular assembly for high-performance PRET.
  • To achieve NIR delayed fluorescence emission for enhanced bioimaging capabilities.
  • To demonstrate the system's potential in live-cell imaging and other applications.

Main Methods:

  • Construction of a tailored charge transfer supramolecular assembly using dibenzyl-bridged pyridinium derivative (G1), cucurbit[8]uril (CB[8]), and β-cyclodextrin-grafted hyaluronic acid (HACD).
  • Investigation of the assembly's structural transformation and energy transfer dynamics.
  • Characterization of PRET from bromophenyl pyridinium (donor) to anthracenyl pyridinium (acceptor) with NIR delayed fluorescence emission.
  • Evaluation of the system's performance in live-cell labeling and detection.

Main Results:

  • A supramolecular assembly was successfully constructed, exhibiting a large Stokes shift (320 nm) and efficient energy transfer.
  • The system achieved single-molecule PRET, expanding phosphorescence to 650 nm NIR delayed fluorescence with a lifetime of 11.20 µs.
  • Successful NIR channel labeling and detection in living cells were demonstrated, validating its bioimaging potential.

Conclusions:

  • The developed supramolecular PRET system overcomes traditional wavelength limitations, enabling high-performance NIR bioimaging.
  • Rational structural tailoring and confinement engineering are key to advancing supramolecular PRET design.
  • This work holds broad significance for cell imaging, information encryption, and anti-counterfeiting applications.