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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.
Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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 12, 2026

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

Rapid-scanning frequency-domain fluorometer with picosecond time resolution.

F V Bright, G M Hieftje

    Applied Optics
    |May 22, 2010
    PubMed
    Summary
    This summary is machine-generated.

    A novel frequency-domain fluorometer operates from 10 MHz to 1 GHz, enabling rapid excited-state lifetime measurements. This instrument achieves high signal-to-noise ratios and precise decay determinations for advanced fluorescence spectroscopy.

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    Published on: June 27, 2014

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    Last Updated: Jun 12, 2026

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
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    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

    Published on: January 18, 2017

    Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
    10:03

    Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy

    Published on: June 27, 2014

    Area of Science:

    • Spectroscopy
    • Physical Chemistry
    • Instrumentation

    Background:

    • Frequency-domain fluorometry is crucial for analyzing excited-state dynamics.
    • Existing instruments may have limitations in speed, frequency range, or precision.
    • Accurate determination of fluorescence lifetimes is essential in various scientific fields.

    Purpose of the Study:

    • To introduce a new frequency-domain fluorometer with enhanced capabilities.
    • To demonstrate the instrument's performance in terms of speed, signal-to-noise ratio, and precision.
    • To enable the resolution of complex fluorescence decay behaviors.

    Main Methods:

    • Utilized a mode-locked Ar-ion laser as an excitation source.
    • Employed an extracavity acoustooptic light modulator for frequency generation.
    • Developed a rapid data acquisition system for spectral analysis.

    Main Results:

    • The fluorometer operates across a 10 MHz to 1 GHz frequency range.
    • Acquisition of full frequency spectra in approximately 10 seconds.
    • Achieved signal-to-noise ratios of at least 20 for moderate fluorescence signals.
    • Demonstrated the capability to resolve multiple exponential decays and determine single exponential decays with <20 ps precision.

    Conclusions:

    • The new frequency-domain fluorometer offers a rapid and precise method for fluorescence lifetime analysis.
    • The instrument's performance characteristics make it suitable for detailed studies of excited-state dynamics.
    • This advancement facilitates more accurate and efficient fluorescence spectroscopy research.