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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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Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview01:13

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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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Super-resolution Fluorescence Microscopy01:37

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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: Apr 4, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
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Total Internal Reflection Fluorescence Microscopy.

Ahmet Yildiz, Ronald D Vale

    Cold Spring Harbor Protocols
    |September 3, 2015
    PubMed
    Summary
    This summary is machine-generated.

    Total internal reflection fluorescence microscopy (TIRFM) offers enhanced sensitivity for biological imaging by reducing background noise. This technique illuminates molecules near the coverslip, enabling detailed study of cell membrane interactions in living cells.

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    Area of Science:

    • Biophysics
    • Cell Biology
    • Microscopy

    Background:

    • Epifluorescence microscopy struggles with weak signals due to out-of-focus light.
    • Confocal microscopy improves optical sectioning but sacrifices some in-focus photons.
    • Total internal reflection fluorescence microscopy (TIRFM) addresses these limitations.

    Purpose of the Study:

    • To highlight the advantages of TIRFM in fluorescence microscopy.
    • To explain the principles and applications of TIRFM in biological imaging.
    • To emphasize TIRFM's utility for studying cell membrane dynamics.

    Main Methods:

    • Utilizes a wide-field illumination technique.
    • Restricts illumination to the region near the glass-water interface.
    • Employs total internal reflection to minimize background signal.

    Main Results:

    • Achieves significantly reduced background noise.
    • Provides high temporal resolution for dynamic processes.
    • Enables sensitive detection of fluorescent signals.

    Conclusions:

    • TIRFM is a powerful tool for sensitive detection of fluorescently labeled molecules.
    • Its ability to image near the cell membrane makes it ideal for studying molecular interactions.
    • TIRFM facilitates research in areas like protein studies, signaling cascades, and cell dynamics.