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Related Concept Videos

Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

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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...
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Total Internal Reflection Fluorescence Microscopy01:05

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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.
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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...
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Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
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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.
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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
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Applications of imaging fluorescence correlation spectroscopy.

Anand P Singh1, Thorsten Wohland1

  • 1Department of Biological Sciences and Centre for Bio-Imaging Sciences, National University of Singapore, Singapore, 117557, Singapore; Department of Chemistry and Centre for Bio-Imaging Sciences, National University of Singapore, Singapore, 117557, Singapore.

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Imaging fluorescence correlation spectroscopy (imaging FCS) offers high spatial and temporal resolution for quantitative bioimaging. This review highlights recent applications of this advanced technique in biological research.

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Area of Science:

  • Quantitative bioimaging
  • Biophysical techniques

Background:

  • Imaging fluorescence correlation spectroscopy (imaging FCS) is an emerging quantitative bioimaging technique.
  • It enables the acquisition of fluorescence correlation functions across contiguous points in an image format.
  • This method offers high time resolution and single-molecule sensitivity.

Purpose of the Study:

  • To review the applications of imaging FCS in biological problems over the last two years.
  • To provide an overview of the capabilities of imaging FCS techniques.

Main Methods:

  • Implementation of imaging FCS across various modalities.
  • Combination with super-resolution techniques to achieve high spatial and temporal resolution.

Main Results:

  • Demonstration of imaging FCS's utility in diverse biological applications.
  • Highlighting the technique's capacity for simultaneous high spatial and temporal resolution.

Conclusions:

  • Imaging FCS is a powerful tool for quantitative bioimaging.
  • Its recent applications showcase its growing impact on biological research.