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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.
Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

Atomic fluorescence spectroscopy (AFS) is an analytical technique that involves the electronic transitions of atoms in a flame, furnace, or plasma being excited by electromagnetic (EM) radiation. When these atoms absorb energy, they become excited and subsequently release energy as they return to their original state. This emitted light, or "fluorescence," is observed at a right angle to the incident beam. Both absorption and emission processes transpire at distinct wavelengths, which are...
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 5, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
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Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

EMCCD-based spectrally resolved fluorescence correlation spectroscopy.

Felix Bestvater1, Zahir Seghiri, Moon Sik Kang

  • 1Cell Biophysics Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea.

Optics Express
|December 18, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces spectrally resolved fluorescence correlation spectroscopy (FCS) for enhanced fluorophore analysis. The new method allows for detailed characterization of photophysical properties and improved intracellular measurements.

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Last Updated: Jun 5, 2026

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Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)
10:59

Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)

Published on: April 6, 2012

Area of Science:

  • Biophysics
  • Spectroscopy
  • Microscopy

Background:

  • Fluorescence Correlation Spectroscopy (FCS) is a powerful technique for studying molecular dynamics.
  • Conventional FCS setups often lack the spectral resolution to differentiate complex fluorophore mixtures.
  • Advanced detection methods are needed to enhance the capabilities of FCS.

Purpose of the Study:

  • To implement and validate a novel spectrally resolved detection scheme for FCS.
  • To demonstrate the capability of this system for analyzing fluorophore photophysical properties.
  • To assess the benefits of spectral flexibility in optimizing FCS experiments.

Main Methods:

  • Integration of a prism-based spectrometer and an electron-multiplying charge-coupled device (EMCCD) camera into a confocal laser scanning/FCS microscope.
  • High-speed spectral data acquisition (over 80,000 spectra/second) enabling single photon counting.
  • In vitro characterization of spectrally distinct quantum dots and measurement of fluorescence intermittence.

Main Results:

  • Successful identification of up to four spectrally distinct quantum dots in vitro.
  • Demonstration of characterizing photophysical properties by measuring spectral dependence of fluorescence intermittence.
  • Confirmation of intracellular cross-correlation results and validation of spectral flexibility for optimizing detection.

Conclusions:

  • Spectrally resolved detection significantly enhances the capabilities of FCS.
  • This method allows for detailed photophysical characterization and improved analysis of complex biological systems.
  • The developed system offers greater spectral flexibility for optimizing FCS experiments.