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Colloids and Suspensions01:17

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Children at play often make suspensions such as mixtures of mud and water, flour and water, or a suspension of solid pigments in water known as tempera paint. These suspensions are heterogeneous mixtures composed of relatively large particles visible to the naked eye or seen with a magnifying glass. They are cloudy, and the suspended particles settle out after mixing. The suspended particles in a suspension settle out after some time of mixing. The separation of particles from a suspension is...
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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Hyperspectral Imaging as a Tool to Study Optical Anisotropy in Lanthanide-Based Molecular Single Crystals
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[Tyndall's hypochromism in suspensions].

N L Vekshin, M S Frolova, V I Kovalev

    Biofizika
    |April 15, 2015
    PubMed
    Summary

    Scattering in suspensions causes hypochromism, decreasing extinction coefficients. A new method accurately measures true extinction coefficients from diluted suspension spectra, correcting for particle size and refractive index effects.

    Area of Science:

    • Spectroscopy
    • Biophysics
    • Photochemistry

    Context:

    • Light interacts with particles in suspensions through absorption and scattering.
    • Particle size and refractive index influence scattering, leading to hypochromism and reduced extinction coefficients.
    • Tyndall scattering in larger particles is wavelength-independent, causing uniform spectral attenuation.

    Purpose:

    • To introduce a simple method for determining accurate extinction coefficients from absorption or excitation spectra.
    • To correct for scattering-induced hypochromism in diluted suspensions.
    • To analyze spectral data from biological samples like hemoglobin, actinomycin, and flavins.

    Summary:

    • Hypochromism, a decrease in extinction coefficient, arises from light absorption and scattering by particles in suspensions.

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  • This scattering effect is dependent on particle size and refractive index, and minimally on wavelength for larger particles.
  • The study proposes a straightforward technique to derive true extinction coefficients from diluted suspension spectra, mitigating scattering interference.
  • Experimental validation is provided using hemoglobin in erythrocytes, actinomycin in DNA, and flavins in mitochondria.

    • Impact:

    • Enables more accurate spectrophotometric analysis of biological molecules in complex matrices.
    • Provides a crucial correction factor for studies involving light scattering in suspensions.
    • Improves the reliability of quantitative spectral measurements in biophysical and biochemical research.