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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
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Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
09:14

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Published on: October 2, 2012

Absorption effects on total-internal-reflection fluorescence spectroscopy.

M Toriumi, M Yanagimachi, H Masuhara

    Applied Optics
    |August 25, 2010
    PubMed
    Summary
    This summary is machine-generated.

    This study derives a fluorescence intensity formula for optically absorbing samples in total-internal-reflection fluorescence spectroscopy. The findings reveal high depth profile resolution is achievable for absorbing samples.

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    Area of Science:

    • Optics and Photonics
    • Spectroscopy
    • Materials Science

    Background:

    • Total-internal-reflection fluorescence (TIR-F) spectroscopy is a powerful technique for surface analysis.
    • Understanding fluorescence intensity in optically absorbing samples is crucial for accurate depth profiling.

    Purpose of the Study:

    • To theoretically derive a fluorescence intensity formula for optically absorbing samples in TIR-F spectroscopy.
    • To experimentally validate the derived formula and explore its implications for depth profiling.

    Main Methods:

    • Theoretical derivation of fluorescence intensity considering transverse-electric polarization vectors.
    • Mathematical formulation as a function of absorption index and angles.
    • Experimental validation of the derived equations.

    Main Results:

    • A fluorescence intensity formula was successfully derived for absorbing samples.
    • The angular spectrum of fluorescence intensity was shown to be the Laplace transform of effective penetration depth.
    • Experimental confirmation of the theoretical model.

    Conclusions:

    • The developed model accurately describes fluorescence intensity in absorbing samples.
    • High-resolution depth profiling is feasible for absorbing samples using TIR-F spectroscopy.
    • The findings advance the application of TIR-F spectroscopy for complex sample analysis.