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Related Concept Videos

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.
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,...

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Related Experiment Video

Updated: Jun 30, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

High-resolution image-projection fluorescence lifetime imaging microscopy.

WoongJae Baek, Jongchan Park, Liang Gao

    Biorxiv : the Preprint Server for Biology
    |June 29, 2026
    PubMed
    Summary
    This summary is machine-generated.

    Image-projection fluorescence lifetime imaging microscopy (IP-FLIM) enhances speed and resolution by using computational reconstruction. This novel method improves contrast-to-noise ratio and reduces background noise for advanced molecular imaging.

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    Last Updated: Jun 30, 2026

    Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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    Published on: February 9, 2012

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    06:01

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    Published on: April 7, 2023

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
    09:30

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

    Published on: January 18, 2017

    Area of Science:

    • Biophotonics
    • Microscopy
    • Computational Imaging

    Background:

    • Fluorescence lifetime imaging microscopy (FLIM) offers concentration-independent molecular contrast.
    • FLIM faces limitations in acquisition speed, photon dose, and detector complexity.

    Purpose of the Study:

    • To develop an integrated optical and computational platform, image-projection fluorescence lifetime imaging microscopy (IP-FLIM).
    • To overcome the trade-offs in conventional FLIM for high-resolution, component-resolved lifetime imaging.

    Main Methods:

    • Utilized a linear single-photon avalanche diode array for data acquisition.
    • Employed wide-field projection acquisition combined with computational k-space reconstruction.
    • Validated the technique using fluorescent microbeads and endothelial cells.

    Main Results:

    • Achieved high-resolution, component-resolved lifetime imaging.
    • Demonstrated up to 22.3x improvement in contrast-to-noise ratio.
    • Showcased a 72.3% reduction in background noise compared to filtered back-projection.

    Conclusions:

    • IP-FLIM provides a scalable solution for fast, high-resolution multiplex lifetime imaging.
    • The platform integrates optical projection with computational reconstruction for enhanced performance.
    • IP-FLIM addresses key limitations of conventional FLIM techniques.