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Related Concept Videos

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.
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.
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,...
Inductively Coupled Plasma Atomic Emission Spectroscopy: Instrumentation01:26

Inductively Coupled Plasma Atomic Emission Spectroscopy: Instrumentation

Inductively coupled plasma (ICP) is the common plasma source used in atomic emission spectroscopy (AES), a technique that detects and analyzes various elements in a sample. This method is often called inductively coupled plasma atomic emission spectroscopy (ICP-AES).
There are three main types of inductively coupled plasma atomic emission spectroscopy  (ICP-AES) instruments: sequential, simultaneous multichannel, and Fourier transform instruments, with the latter being less commonly used.
Photoluminescence: Fluorescence and Phosphorescence01:23

Photoluminescence: Fluorescence and Phosphorescence

Photoluminescence is a process where a molecule absorbs light energy and re-emits it in the form of light. This phenomenon occurs when a substance absorbs photons, promoting its electrons to higher energy level excited states, followed by a relaxation process in which the electrons return to their original ground state energy levels and emit light. Photoluminescence is widely observed in various materials, including semiconductors, and organic and inorganic compounds.
A pair of electrons in a...
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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Related Experiment Video

Updated: Jun 22, 2026

Determination of the Excitation and Coupling Rates Between Light Emitters and Surface Plasmon Polaritons
07:39

Determination of the Excitation and Coupling Rates Between Light Emitters and Surface Plasmon Polaritons

Published on: July 21, 2018

Fluorescence correlation spectroscopy in surface plasmon coupled emission microscope.

J Borejdo, N Calander, Z Gryczynski

    Optics Express
    |June 17, 2009
    PubMed
    Summary
    This summary is machine-generated.

    Surface Plasmon Coupled Emission (SPCE) combined with Fluorescence Correlation Spectroscopy (FCS) offers a powerful method for single-molecule dynamics studies. This technique significantly enhances detection efficiency and reduces background noise for precise molecular analysis.

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

    Published on: December 11, 2021

    Area of Science:

    • Biophysics
    • Spectroscopy
    • Nanotechnology

    Background:

    • Single-molecule dynamics studies require high photon detection rates and low background.
    • Traditional Fluorescence Correlation Spectroscopy (FCS) faces limitations in achieving optimal detection parameters.
    • Enhanced sensitivity is crucial for analyzing molecular behavior at the nanoscale.

    Purpose of the Study:

    • To investigate the application of Surface Plasmon Coupled Emission (SPCE) combined with FCS for improved single-molecule dynamics analysis.
    • To demonstrate the effectiveness of SPCE-FCS in enhancing detection efficiency and reducing background.
    • To characterize the detection volume dimensions achieved by the SPCE-FCS method.

    Main Methods:

    • Utilizing a thin metal film to create a shallow observational volume via evanescent waves generated at the Surface Plasmon Resonance (SPR) angle.
    • Employing metal quenching to further reduce the detection volume to below 10 nm.
    • Collecting emitted fluorescence through the metal film at a specific SPCE angle and using a confocal aperture to define the detection volume.

    Main Results:

    • SPCE-FCS demonstrates high efficiency in measuring molecular diffusion.
    • The detection volume height was precisely determined to be approximately 35 nm on a gold surface.
    • The method effectively meets the stringent requirements for single-molecule dynamics studies.

    Conclusions:

    • SPCE-FCS is a powerful and efficient technique for studying single-molecule dynamics.
    • The shallow detection volume and enhanced signal achieved by SPCE-FCS overcome limitations of conventional methods.
    • This approach offers significant potential for advancing research in molecular interactions and diffusion.