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BO21 exhibits unique fluorescence properties, showing high polarization due to its short lifetime. Its fluorescence changes significantly with heparin and viscosity, suggesting potential as a biosensor for heparin levels and intracellular viscosity.

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Area of Science:

  • Biophysical Chemistry
  • Fluorescence Spectroscopy
  • Molecular Probes

Background:

  • Understanding the fluorescence properties of novel molecular probes is crucial for developing advanced sensing technologies.
  • BO21's structural similarity to fluorescein, yet distinct fluorescence characteristics, warrants detailed investigation.

Purpose of the Study:

  • To investigate the fluorescence properties of BO21 and their dependence on intracellular conditions.
  • To explore the potential of BO21 as a sensor for heparin and intracellular viscosity.

Main Methods:

  • Spectroscopic analysis of BO21 in cell-free solutions under varying pH, temperature, viscosity, and in the presence of proteins and heparin.
  • Measurement of quantum yield, fluorescence polarization (FP), and fluorescence lifetime (FLT).
  • Analytical and experimental investigations, including theoretical calculations.

Main Results:

  • Fluorescence spectra of BO21 are minimally affected by pH and temperature but significantly influenced by viscosity, proteins, and heparin.
  • Heparin induces a red shift in the emission spectrum and increases fluorescence lifetime, suggesting aggregation.
  • BO21 exhibits a surprisingly high FP (0.470) in water due to a very short lifetime, which decreases upon heparin addition.

Conclusions:

  • BO21's high fluorescence polarization is linked to its short fluorescence lifetime.
  • The molecule behaves as a molecular rotor, with its properties modulated by environmental factors like viscosity and heparin.
  • BO21 shows promise as a sensor for detecting heparin levels and intracellular viscosity.