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Related Concept Videos

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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Related Experiment Video

Updated: Jun 16, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Two-chopper technique for measuring fluorescence lifetimes.

K A Ingersoll

    Applied Optics
    |February 16, 2010
    PubMed
    Summary

    This study introduces a two chopper technique for accurate fluorescence lifetime measurements. This method avoids relaxation delays and detector saturation issues, improving experimental efficiency.

    Area of Science:

    • Spectroscopy
    • Photophysics
    • Analytical Chemistry

    Background:

    • Fluorescence lifetime measurements are crucial for understanding molecular dynamics.
    • Direct excitation into the fluorescing level can lead to detector saturation.
    • Relaxation delays from higher pump bands complicate lifetime analysis.

    Purpose of the Study:

    • To present a novel two chopper technique for fluorescence lifetime measurements.
    • To address challenges associated with direct excitation at the same wavelength as fluorescence.
    • To improve accuracy and simplify fluorescence lifetime determination.

    Main Methods:

    • Utilizing a two chopper system to control pump and detection.
    • Implementing direct pumping into the emitting state.

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  • Measuring fluorescence lifetimes under specific experimental conditions.

    • Main Results:

    • Successfully measured fluorescence lifetimes with the two chopper technique.
    • Avoided delays caused by relaxation from higher energy states.
    • Prevented detector saturation issues during measurements.

    Conclusions:

    • The two chopper technique offers a robust method for fluorescence lifetime analysis.
    • This technique simplifies experiments by avoiding indirect excitation pathways.
    • It enhances the reliability of fluorescence lifetime data, particularly in challenging scenarios.