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Caffeine interaction with fluorescent calcium indicator dyes.

M Muschol1, B R Dasgupta, B M Salzberg

  • 1Department of Neuroscience, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6074 USA.

Biophysical Journal
|July 2, 1999
PubMed
Summary
This summary is machine-generated.

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Caffeine significantly alters the fluorescence of common calcium indicator dyes, affecting measurements of intracellular calcium. This interaction impacts dyes like mag-fura-2 and fluo-3, potentially distorting scientific results.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Pharmacology

Background:

  • Calcium indicator dyes are crucial for measuring intracellular calcium concentrations.
  • Caffeine is a widely used pharmacological agent with potential interactions in biological assays.

Purpose of the Study:

  • To investigate the interaction between caffeine and common calcium indicator dyes.
  • To determine if caffeine affects the fluorescence properties of these dyes.
  • To assess the impact of these interactions on the accuracy of intracellular ion measurements.

Main Methods:

  • Tested interactions of caffeine (millimolar concentrations) with mag-fura-2, magnesium green, fura-2, and fluo-3.
  • Analyzed changes in fluorescence intensity and spectra.
  • Examined effects on calcium binding affinities.

Related Experiment Videos

  • Investigated interactions with non-chelating fluorescent dyes (fluorescein, sulforhodamine 101).
  • Varied solution hydrophobicity using water/dioxane mixtures.

    • Main Results:

    • Caffeine significantly altered fluorescence intensities of tested calcium indicator dyes (enhancement or diminution).
    • Caffeine-induced spectral changes differed from metal ion binding.
    • Slight increases in calcium binding affinity were observed for mag-fura-2 and magnesium green.
    • Caffeine affected fluorescence of non-chelating dyes, indicating interaction with the fluorophore.
    • Hydrophobicity variations mimicked caffeine's effects, suggesting a general hydrophobic interaction.

    Conclusions:

    • Caffeine interacts directly with the fluorophore of calcium indicator dyes.
    • These interactions can significantly distort fluorescence measurements of intracellular calcium.
    • Hydrophobic substances generally can alter dye fluorescence in a dye-specific manner.
    • Caution is advised when using caffeine or similar agents in fluorescence-based ion concentration studies.