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Lifetime-based pH sensors: indicators for acidic environments.

H J Lin1, H Szmacinski, J R Lakowicz

  • 1Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 21201, USA.

Analytical Biochemistry
|March 30, 1999
PubMed
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We identified new lifetime-based pH sensors, Oregon green 514 carboxylic acid, Cl-NERF, and DM-NERF, which are effective in acidic environments. These probes offer precise pH monitoring in cellular compartments.

Area of Science:

  • Analytical Chemistry
  • Biophysical Chemistry
  • Molecular Probes

Background:

  • Fluorescence-based pH sensing is crucial for biological research.
  • Existing sensors often lack sensitivity in acidic cellular compartments.
  • Oregon green 514 carboxylic acid, Cl-NERF, and DM-NERF are photostable, high quantum yield probes.

Purpose of the Study:

  • To characterize pH-dependent fluorescence intensity decays of three fluorophores.
  • To identify novel lifetime-based sensors for low pH environments.
  • To evaluate the suitability of these probes for monitoring acidic cellular compartments.

Main Methods:

  • Frequency-domain fluorometry was employed to analyze fluorescence intensity decays.
  • Global intensity decay analysis was performed across a range of pH values.

Related Experiment Videos

  • Titration curves of phase angles and modulations versus pH were used to determine pKa values.

    • Main Results:

    • Fluorescence intensity decays of the probes were highly pH-dependent.
    • Double exponential decays were observed at intermediate pH, with distinct lifetimes for protonated and unprotonated forms.
    • Apparent pKa values ranged from pH 3 to 5, indicating suitability for acidic environments.

    Conclusions:

    • Oregon green 514 carboxylic acid, Cl-NERF, and DM-NERF function as effective lifetime-based pH sensors.
    • These probes exhibit unique characteristics for monitoring pH in acidic cellular compartments.
    • The identified pKa range fills a gap in available lifetime-based pH sensing technologies.