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Related Concept Videos

Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

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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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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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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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Atomic Fluorescence Spectroscopy01:29

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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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Variables Affecting Phosphorescence and Fluorescence01:26

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Fluorescence and phosphorescence are essential phenomena in fields like analytical chemistry, biological imaging, and materials science, where they detect molecular properties and visualize cellular structures. Understanding the variables that influence these luminescent behaviors is crucial for maximizing accuracy and efficiency in their applications. These variables can broadly be grouped into chemical structure, solvent properties, and external conditions, each playing a distinct role in...
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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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Updated: Aug 26, 2025

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

Juan Jeremías Incicco1,2, Debjit Roy1,2, Melissa D Stuchell-Brereton1,2

  • 1Department of Biochemistry and Molecular Biophysics, Washington University in St Louis, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 13, 2022
PubMed
Summary
This summary is machine-generated.

Fluorescence Correlation Spectroscopy (FCS) helps identify phase boundaries for biomolecular condensate formation. This method also reveals crucial transport properties within these condensates, advancing our understanding of their assembly and function.

Keywords:
Biomolecular condensatesFluorescence Correlation SpectroscopyIntrinsically disordered proteinsPhase separation

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Chemical Physics

Background:

  • Biomolecular condensates are essential for cellular organization and function.
  • Understanding their formation requires identifying phase boundaries and tie-lines.
  • Quantitative insights into condensate assembly and function are needed.

Purpose of the Study:

  • To present Fluorescence Correlation Spectroscopy (FCS) as a versatile method for studying biomolecular condensates.
  • To demonstrate FCS's capability in estimating phase boundaries for various solutions.
  • To highlight FCS's utility in characterizing condensate transport properties.

Main Methods:

  • Utilizing Fluorescence Correlation Spectroscopy (FCS) for quantitative analysis.
  • Applying FCS to single-component and multicomponent solutions.
  • Analyzing condensate formation and properties through spectroscopic measurements.

Main Results:

  • FCS effectively estimates phase boundaries for biomolecular condensate formation.
  • The technique provides insights into the transport properties within condensates.
  • Successful application demonstrated for both single- and multicomponent systems.

Conclusions:

  • Fluorescence Correlation Spectroscopy is a powerful tool for characterizing biomolecular condensate phase behavior.
  • FCS enables a quantitative understanding of the forces governing condensate assembly.
  • This approach offers valuable insights into the dynamic properties of condensates.