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Related Concept Videos

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.

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Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
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Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Published on: August 2, 2018

Electron multiplying CCD based detection for spatially resolved fluorescence correlation spectroscopy.

Markus Burkhardt, Petra Schwille

    Optics Express
    |June 12, 2009
    PubMed
    Summary
    This summary is machine-generated.

    Electron multiplying CCD (EMCCD) cameras enable spatially resolved fluorescence correlation spectroscopy (FCS). This advanced method achieves high time resolution, matching standard detectors for precise diffusion measurements.

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    Last Updated: Jun 22, 2026

    Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy
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    Published on: August 2, 2018

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    08:32

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    12:51

    Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

    Published on: December 9, 2013

    Area of Science:

    • Biophysics
    • Optical Microscopy
    • Spectroscopy

    Background:

    • Fluorescence correlation spectroscopy (FCS) is a powerful technique for studying molecular dynamics.
    • Traditional FCS detection often relies on point detectors like avalanche photodiodes.
    • Electron multiplying CCD (EMCCD) cameras offer high sensitivity and rapid readout capabilities.

    Purpose of the Study:

    • To evaluate the suitability of EMCCD cameras for performing fluorescence correlation spectroscopy.
    • To compare EMCCD-based FCS with standard avalanche photodiode detection.
    • To explore the potential of EMCCD cameras for spatially resolved FCS applications.

    Main Methods:

    • Implementing an EMCCD camera for FCS measurements.
    • Comparing autocorrelation curves obtained with EMCCD and avalanche photodiode detectors.
    • Utilizing different EMCCD readout modes to achieve high time resolution (down to 20 microseconds).
    • Performing proof-of-principle FCS measurements with multiple excitation volumes.

    Main Results:

    • EMCCD-based FCS demonstrated good agreement with standard avalanche photodiode detection.
    • Achieved time resolution was sufficient to resolve the diffusion of free dye molecules.
    • Flexible area detection with EMCCD cameras was advantageous for spatially resolved FCS.
    • Successful demonstration of FCS with two excitation volumes using EMCCD detection.

    Conclusions:

    • EMCCD cameras are a viable and effective alternative for FCS measurements.
    • The high time resolution and flexible detection capabilities of EMCCDs expand FCS applications, particularly for spatially resolved studies.
    • EMCCD-based FCS can be integrated into wide-field imaging and multi-spot confocal microscopy for enhanced spatial analysis.