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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.
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400 keV in...

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Related Experiment Video

Updated: Jun 12, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

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Published on: May 3, 2022

Photon tunneling microscopy.

J M Guerra

    Applied Optics
    |June 23, 2010
    PubMed
    Summary
    This summary is machine-generated.

    Researchers developed a novel photon tunneling microscopy technique to image surface topography with nanometer resolution. This method offers enhanced lateral and vertical precision for analyzing micro- and nanostructures.

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    Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

    Published on: June 7, 2019

    Area of Science:

    • Optics and Photonics
    • Surface Science
    • Microscopy

    Background:

    • Total internal reflection (TIR) is a phenomenon used in various optical techniques.
    • Microscopy relies on light interaction with samples to reveal surface details.
    • Submicron gap imaging presents challenges in achieving high resolution.

    Purpose of the Study:

    • To develop a novel method for high-resolution surface topography imaging.
    • To utilize photon tunneling through a submicron gap for quantitative measurements.
    • To demonstrate the capabilities of this technique on various microstructures.

    Main Methods:

    • Incident photons tunnel through a submicron gap at a TIR surface, acting as the object plane.
    • Dielectric samples influence tunneling, which increases exponentially with height.
    • Video photometry and a three-axis oscilloscope convert tunneling images into real-time 3-D topographic maps.

    Main Results:

    • Achieved vertical resolution below a nanometer and lateral resolution of approximately 0.29 lambda.
    • Demonstrated variable perspective 3-D topographic imaging.
    • Successfully imaged various surfaces including diamond-turned, optical data structures, and microlithographic patterns.

    Conclusions:

    • Photon tunneling microscopy provides a powerful new tool for nanoscale surface metrology.
    • The technique offers high resolution and quantitative 3-D topographic information.
    • It shows promise for analyzing micro- and nanostructures in diverse scientific and engineering fields.