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Updated: Jan 19, 2026

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
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Confocal fluorescence correlation spectroscopy through a sparse layer of scattering objects.

Anirban Sarkar, Joseph Gallagher, Irène Wang

    Optics Express
    |September 11, 2019
    PubMed
    Summary
    This summary is machine-generated.

    Scattering layers bias fluorescence correlation spectroscopy (FCS) measurements of molecular properties. Theoretical modeling explains how bead size and refractive index impact FCS data through diffraction and refraction.

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    Area of Science:

    • Biophysics
    • Optical Microscopy
    • Spectroscopy

    Background:

    • Optical microscopy faces challenges in scattering media like biological tissue.
    • Scattering significantly impacts image quality and quantitative molecular property determination.

    Purpose of the Study:

    • To investigate the effects of light scattering on fluorescence correlation spectroscopy (FCS).
    • To analyze how scattering layers influence quantitative molecular measurements.

    Main Methods:

    • Conducted fluorescence correlation spectroscopy (FCS) experiments.
    • Used a solution of fluorophores and a scattering layer of dielectric beads.
    • Developed theoretical modeling to analyze scattering effects.

    Main Results:

    • Observed a decrease in fluorescence signal with increasing distance from the scattering layer.
    • Found that the estimated molecular number is biased below a characteristic distance (twice bead diameters).
    • Theoretical modeling confirmed the influence of bead size and refractive index on FCS measurements via diffraction and refraction.

    Conclusions:

    • Scattering layers significantly affect FCS measurements, particularly the estimation of molecular numbers.
    • Understanding diffraction and refraction effects is crucial for accurate quantitative analysis in scattering media.
    • The study provides insights into mitigating scattering artifacts in optical measurements.