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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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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Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: May 2, 2026

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
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Axial nanoscale localization by normalized total internal reflection fluorescence microscopy.

Marcelina Cardoso Dos Santos, Régis Déturche, Cyrille Vézy

    Optics Letters
    |February 25, 2014
    PubMed
    Summary
    This summary is machine-generated.

    Researchers developed a simple microscope modification for nanometric axial resolution, achieving 10 nm. This technique uses normalized total internal reflection and epi-illumination imaging to study vesicle adhesion.

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    Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy VA-TIRFM
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    Area of Science:

    • Biophysics
    • Microscopy
    • Cell Biology

    Background:

    • Standard total internal reflection fluorescence microscopy lacks high axial resolution.
    • Studying the adhesion dynamics of phospholipid giant unilamellar vesicles requires precise axial measurements.

    Purpose of the Study:

    • To present a straightforward modification to total internal reflection fluorescence microscopy.
    • To achieve nanometric axial resolution for enhanced imaging capabilities.
    • To demonstrate the application of this technique in studying vesicle adhesion.

    Main Methods:

    • Modification of a standard total internal reflection fluorescence microscope.
    • Normalization of total internal reflection images with conventional epi-illumination images.
    • Application of the developed method to analyze phospholipid giant unilamellar vesicle adhesion.

    Main Results:

    • Achieved nanometric axial resolution, typically around 10 nm.
    • Successfully demonstrated the technique's potential for studying vesicle adhesion.
    • Provided a new method for high-resolution imaging in biophysical studies.

    Conclusions:

    • The presented microscope modification offers a simple yet effective way to enhance axial resolution.
    • This technique is valuable for investigating the adhesion mechanisms of biological vesicles.
    • The method opens new avenues for high-precision imaging in cell biology and biophysics.