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Related Experiment Video

Updated: Jul 9, 2026

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
08:27

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

A high-precision ratiometric fluorosensor for pH: implementing time-dependent non-linear calibration protocols for

Aron Hakonen1, Stefan Hulth

  • 1Department of Chemistry, GöteborgUniversity, SE-412 96 Göteborg, Sweden. hakonen@chem.gu.se

Analytica Chimica Acta
|December 11, 2007
PubMed
Summary

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Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

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A new calibration protocol enhances optical pH sensor performance using immobilized 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS). This method improves sensor accuracy, precision, and lifetime by compensating for calibration drift and reducing photoacidity.

Area of Science:

  • Analytical Chemistry
  • Chemical Sensors
  • Materials Science

Background:

  • Optical sensors offer advantages for pH monitoring.
  • Ratiometric fluorophores like HPTS are sensitive to pH changes.
  • Immobilization strategies can enhance sensor stability and performance.

Purpose of the Study:

  • To develop and validate a time-dependent, non-linear calibration protocol for optical pH sensors.
  • To improve the precision, accuracy, and operational lifetime of HPTS-based optical sensors.
  • To investigate the impact of immobilization on HPTS photoacidity and excited-state dynamics.

Main Methods:

  • Implemented a versatile time-dependent non-linear calibration protocol.
  • Utilized 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) immobilized in ethyl-cellulose.

Related Experiment Videos

Last Updated: Jul 9, 2026

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
08:27

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

  • Employed a dual excitation/dual emission ratiometric sensing scheme (405/440 nm and 465/510 nm).

    • Main Results:

    • The protocol compensated for progressive calibration parameter drift, enhancing sensor precision and accuracy.
    • Significantly reduced photoacidity of immobilized HPTS was observed and utilized.
    • The dual excitation/dual emission scheme amplified sensor response, improving signal-to-noise ratio by ~400% compared to single emission.
    • Achieved excellent repeatability (0.0044 pH units), comparable to the standardization electrode.

    Conclusions:

    • The developed calibration protocol significantly enhances the performance of optical pH sensors.
    • Immobilization of HPTS in ethyl-cellulose offers reduced photoacidity and improved sensor characteristics.
    • The optimized ratiometric sensing scheme provides superior signal enhancement for pH monitoring.