Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Two-Dimensional Microscopy in Microbiology

Two-Dimensional Microscopy in Microbiology

Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...

Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

Two-Dimensional (2D) NMR: Overview

Two-Dimensional (2D) NMR: Overview

The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse.

2D NMR: Overview of Homonuclear Correlation Techniques

2D NMR: Overview of Homonuclear Correlation Techniques

Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...

Fluorescence and Phosphorescence: Instrumentation

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.

Confocal Fluorescence Microscopy

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,...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Structural Transformation of Hydrated Excess Protons at the Water Surface.

Journal of the American Chemical Society·2026

Same author

Distinct adsorption behavior and structures of cell-penetrating peptides at a model lipid membrane interface: a heterodyne-detected vibrational sum frequency generation spectroscopy study.

Chemical science·2026

Same author

Inhomogeneity and Vibrational Coupling of Free OH Stretch at the Air/Water Interface.

Journal of the American Chemical Society·2026

Same author

Localization of the Au-Au bond strength in the triplet excited state of the [Au(CN)<sub>2</sub> <sup>-</sup>] oligomers revealed by ultrafast time-domain Raman spectroscopy.

Chemical science·2025

Same author

Probing Protein-DNA Interactions with Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy Utilizing Protein-Induced Fluorescence Enhancement.

The journal of physical chemistry. B·2025

Same author

Development of a setup for interferometric two-dimensional heterodyne-detected vibrational sum-frequency generation (2D HD-VSFG) measurements to study liquid interfaces with sub-100 fs time resolution.

The Journal of chemical physics·2025

Search research articles

Related Experiment Video

Updated: May 8, 2026

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

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

Published on: December 11, 2021

Two-dimensional fluorescence lifetime correlation spectroscopy. 2. Application.

Kunihiko Ishii¹, Tahei Tahara

¹Molecular Spectroscopy Laboratory, RIKEN , 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

The Journal of Physical Chemistry. B

|August 28, 2013

Summary

This summary is machine-generated.

This study experimentally implements two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS). The method reveals microsecond dynamics of biological molecules, like DNA hairpin structures, by analyzing unique fluorescence lifetime distributions.

More Related Videos

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

Related Experiment Videos

Last Updated: May 8, 2026

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

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

Published on: December 11, 2021

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

Area of Science:

Biophysics
Spectroscopy
Molecular Dynamics

Background:

Two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS) provides a theoretical framework for analyzing molecular dynamics.
Experimental implementation is crucial for validating theoretical models and applying them to real-world systems.

Purpose of the Study:

To experimentally implement and validate two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS).
To demonstrate the capability of 2D FLCS in analyzing molecular dynamics and species identification in complex systems.

Main Methods:

Utilizing time-correlated single photon counting (TCSPC) to acquire photon data.
Constructing two-dimensional emission-delay correlation maps.
Applying the maximum entropy method (MEM) for reliable inverse Laplace transformation to obtain 2D lifetime correlation maps.

Main Results:

Successfully applied 2D FLCS to a dye mixture, identifying different species without prior knowledge.
Demonstrated 2D FLCS's ability to visualize microsecond dynamics of a DNA hairpin by distinguishing species via unique lifetime distributions.
Observed equilibrium dynamics between open and closed DNA hairpin forms through cross-peaks in 2D correlation maps.

Conclusions:

The developed 2D FLCS is experimentally feasible and effective for species identification in inhomogeneous samples.
2D FLCS offers a powerful tool for visualizing and understanding the spontaneous structural dynamics of biological molecules with microsecond resolution.