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Light scattering and fluorescence by small particles having internal structure.

M Kerker, H Chew, P J McNulty

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |January 1, 1979
    PubMed
    Summary
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    Light scattering signals reveal particle internal morphology in backward directions. Fluorescence signals depend on molecule distribution within small particles, impacting quantitative analysis.

    Area of Science:

    • Optical physics
    • Biophysics
    • Computational modeling

    Background:

    • Elastic light scattering and fluorescence are crucial for analyzing small particles, particularly biological cells.
    • Internal particle morphology and molecule distribution can significantly influence optical signals.
    • Previous models often simplify particle properties, potentially leading to inaccurate interpretations.

    Purpose of the Study:

    • To investigate the impact of internal particle morphology on elastic light scattering signals.
    • To determine how the spatial distribution of fluorescent molecules affects fluorescence signals within small particles.
    • To provide insights for improving quantitative analysis using light scattering and fluorescence.

    Main Methods:

    • Development of a computational algorithm for analyzing light scattering from small particles (diameter < 1 micron).

    Related Experiment Videos

  • Modeling of fluorescent molecule distribution within particles and its effect on emission spectra and signals.
  • Calculations restricted to small particles with isotropic and immobile fluorescent molecules.

    • Main Results:

    • Elastic light scattering is sensitive to internal morphology primarily in backward directions for small particles.
    • Fluorescence signals are not simply proportional to molecule concentration; geometrical and optical properties play a role.
    • Morphological features can dramatically alter measured fluorescence, including emission spectra.

    Conclusions:

    • Backward direction observations are recommended for probing particle internal morphology via light scattering.
    • Quantitative fluorescence measurements in small particles must account for molecule distribution and particle properties.
    • These findings can aid in discriminating cells based on morphology or fluorescent moiety distribution and suggest polarization strategies to minimize effects.