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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: Sep 20, 2025

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
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Photonic lantern TIRF microscopy for highly efficient, uniform, artifact-free imaging.

Abdullah Husain, Stephanos Yerolatsitis, Rodrigo Amezcua Correa

    Optics Express
    |November 22, 2024
    PubMed
    Summary

    We developed a photonic lantern TIRF (total internal reflection fluorescence) method for uniform, artifact-free illumination. This technique simplifies TIRF microscopy, enabling unbiased imaging and advanced applications like super-resolution microscopy.

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    Area of Science:

    • Optical Microscopy
    • Biophysics
    • Photonics

    Background:

    • Total internal reflection fluorescence (TIRF) microscopy is crucial for live-cell imaging, but achieving uniform excitation can be challenging.
    • Existing TIRF methods often involve complex setups with moving parts, limiting their practicality and robustness.

    Purpose of the Study:

    • To introduce a novel method for generating uniform, artifact-free TIRF excitation using a photonic lantern.
    • To overcome the limitations of conventional TIRF illumination systems.

    Main Methods:

    • A tapered waveguide photonic lantern with a multimode input and nine few-mode outputs was designed and fabricated.
    • The photonic lantern was utilized for simultaneous multi-beam TIRF illumination from nine azimuthal directions.
    • The system's performance was characterized, including TIRF penetration depth tuning and light source compatibility.

    Main Results:

    • The photonic lantern enabled single-shot, uniform, and artifact-free TIRF excitation without non-stationary devices.
    • The system demonstrated low-loss light transmission, supporting various light sources and wavelengths.
    • High-quality excitation facilitated unbiased imaging across the entire field-of-view.

    Conclusions:

    • The photonic lantern TIRF (PL-TIRF) technique offers a simple, robust, and advantageous alternative to existing TIRF methods.
    • PL-TIRF enables advanced imaging applications, including flat-field super-resolution and shadowless live-cell imaging.