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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.
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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.
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...

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Related Experiment Video

Updated: Jun 22, 2026

Simultaneous Brightfield, Fluorescence, and Optical Coherence Tomographic Imaging of Contracting Cardiac Trabeculae Ex Vivo
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Simultaneous Brightfield, Fluorescence, and Optical Coherence Tomographic Imaging of Contracting Cardiac Trabeculae Ex Vivo

Published on: October 2, 2021

Non-contact fluorescence optical tomography with scanning patterned illumination.

Amit Joshi, Wolfgang Bangerth, Eva M Sevick-Muraca

    Optics Express
    |June 12, 2009
    PubMed
    Summary

    This study introduces a new non-contact fluorescence optical tomography method using multiple illumination patterns to improve imaging of tissue fluorescence. The advanced technique enhances accuracy and detail in reconstructing internal fluorescence distributions.

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    Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy (oSLO) and Optical Coherence Tomography (OCT)

    Published on: August 4, 2018

    Area of Science:

    • Biomedical Optics
    • Medical Imaging
    • Fluorescence Tomography

    Background:

    • Optical tomography faces challenges with ill-posed inverse problems for accurate tissue fluorescence mapping.
    • Existing methods struggle to precisely recover interior fluorescence yield distributions from boundary measurements.

    Purpose of the Study:

    • To develop a novel non-contact fluorescence optical tomography (FOT) scheme.
    • To reduce the ill-posedness of inverse problems in FOT using multiple area illumination patterns.
    • To enhance the accuracy of reconstructing interior fluorescence yield distributions in biological tissues.

    Main Methods:

    • Utilized multiple area illumination patterns (line, Gaussian, diffractive optics) on a simulated tissue phantom.
    • Formulated image reconstruction as an optimization problem minimizing a regularized difference between observed and predicted boundary measurements.
    • Processed multiple datasets simultaneously using a parallel framework for fluorescence tomography.
    • Reconstructed fluorescence distributions for single and multiple targets (5mm diameter) at depths of 1-2cm.

    Main Results:

    • Demonstrated successful image reconstructions of fluorescent targets within a simulated tissue phantom.
    • Achieved qualitative and quantitative improvements compared to reconstructions using single measurement data.
    • Validated the effectiveness of multiple illumination patterns in enhancing FOT accuracy.

    Conclusions:

    • The novel non-contact FOT scheme effectively improves the reconstruction of interior fluorescence distributions.
    • Employing multiple area illumination patterns significantly reduces the ill-posedness inherent in FOT inverse problems.
    • This approach offers enhanced accuracy for biomedical imaging applications.