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A continuously variable frequency cross-correlation phase fluorometer with picosecond resolution.

E Gratton, M Limkeman

    Biophysical Journal
    |December 1, 1983
    PubMed
    Summary
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    This study details a variable frequency cross-correlation phase fluorometer for precise lifetime measurements. The instrument achieves picosecond resolution, enabling accurate analysis of complex decay processes.

    Area of Science:

    • Analytical Chemistry
    • Physical Chemistry
    • Spectroscopy

    Background:

    • Phase fluorometry is a powerful technique for determining molecular lifetimes.
    • Accurate measurements require instruments with high frequency response and resolution.
    • Understanding systematic errors is crucial for reliable data.

    Purpose of the Study:

    • To describe the construction and performance of a novel variable frequency cross-correlation phase fluorometer.
    • To analyze sources of systematic errors affecting phase fluorometer measurements.
    • To demonstrate the instrument's capability in measuring diverse molecular lifetimes.

    Main Methods:

    • Construction of a variable frequency cross-correlation phase fluorometer operating from 1-160 MHz.
    • Analysis of signal distortions from light modulators and dynode characteristics.

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  • Investigation of systematic errors, including nonhomogeneous modulation and photomultiplier color effects.

    • Main Results:

    • The phase fluorometer achieves a resolution of a few picoseconds.
    • Distortion effects and systematic errors were quantified and discussed.
    • Successful application to measuring very long, very short, and multiexponential lifetimes was demonstrated.

    Conclusions:

    • The developed phase fluorometer offers high precision for fluorescence lifetime measurements.
    • The study provides insights into mitigating systematic errors in phase fluorometry.
    • This instrument is suitable for a wide range of applications in photophysics and photochemistry.