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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.
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...
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.
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,...
2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other axis.
2D NMR: Homonuclear Correlation Spectroscopy (COSY)01:06

2D NMR: Homonuclear Correlation Spectroscopy (COSY)

Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...

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Related Experiment Video

Updated: May 11, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

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

Published on: December 11, 2021

Two-photon fluorescence cross-correlation spectroscopy.

P Schwille1, K G Heinze

  • 1Experimental Biophysics Group, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. pschwil@gwdg.de

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry
|May 23, 2013
PubMed
Summary
This summary is machine-generated.

Dual-color cross-correlation spectroscopy analyzes molecular linkages at low concentrations. Two-photon excitation simplifies this technique when using probes with common IR excitation wavelengths.

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Last Updated: May 11, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
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A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
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Area of Science:

  • Biophysics
  • Chemical Physics
  • Spectroscopy

Background:

  • Dual-color cross-correlation spectroscopy (DCCS) is a fluctuation analysis technique.
  • It probes the formation and deletion of linkages between different fluorescently labeled molecules.
  • This method is effective at extremely low molecular concentrations.

Purpose of the Study:

  • To explore the simplification of DCCS using two-photon excitation.
  • To investigate the use of common infrared excitation wavelengths for distinct probe molecules.

Main Methods:

  • Utilizing dual-color cross-correlation spectroscopy.
  • Employing two-photon excitation.
  • Analyzing fluctuation dynamics of fluorescently labeled molecules.

Main Results:

  • Two-photon excitation can simplify DCCS under specific conditions.
  • The simplification is feasible when distinct probe molecules share a common IR excitation wavelength.
  • This approach allows for selective probing of molecular interactions.

Conclusions:

  • Two-photon excitation offers a significant advantage for DCCS.
  • This method enhances the study of molecular interactions at low concentrations.
  • The use of common IR excitation wavelengths broadens the applicability of DCCS.