Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

2.1K
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.
2.1K
Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

1.2K
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...
1.2K
Variables Affecting Phosphorescence and Fluorescence01:26

Variables Affecting Phosphorescence and Fluorescence

3.3K
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...
3.3K
Photoluminescence: Applications01:14

Photoluminescence: Applications

1.3K
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...
1.3K

You might also read

Related Articles

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

Sort by
Same author

Natural killer cells swarm and cross-recruit cytotoxic T cells via CCR5.

Cell reports·2026
Same author

NK cell dysfunction and interferon-γ production underlie autoinflammation in mevalonate kinase deficiency.

Immunity·2026
Same author

Fluorescently Labeled Gradient Hydrogels Reveal Matrix-Dependent Cell Responses to Substrate Stiffness.

Small (Weinheim an der Bergstrasse, Germany)·2026
Same author

Structural organization of p62 filaments and the cellular ultrastructure of calcium-rich p62-enwrapped lipid droplet cargo.

Nature communications·2025
Same author

Macromolecular High-Affinity Binding Probed by Advanced Fluorescence Techniques.

Chembiochem : a European journal of chemical biology·2025
Same author

Correction: Yukhnovets et al. Impact of Molecule Concentration, Diffusion Rates and Surface Passivation on Single-Molecule Fluorescence Studies in Solution. <i>Biomolecules</i> 2022, <i>12</i>, 468.

Biomolecules·2024

Related Experiment Video

Updated: Apr 16, 2026

Absolute Quantum Yield Measurement of Powder Samples
14:20

Absolute Quantum Yield Measurement of Powder Samples

Published on: May 12, 2012

28.9K

Accurate fluorescence quantum yield determination by fluorescence correlation spectroscopy.

Daryan Kempe1, Antonie Schöne2, Jörg Fitter1,2

  • 1†AG Biophysik, I. Physikalisches Institut (IA), RWTH Aachen University, 52062 Aachen, Germany.

The Journal of Physical Chemistry. B
|March 14, 2015
PubMed
Summary
This summary is machine-generated.

This study introduces a fast, accurate fluorescence quantum yield (QY) determination method using fluorescence correlation spectroscopy. It enables photophysical characterization of biomolecules at low concentrations, crucial for single-molecule studies.

More Related Videos

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

11.5K
Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System
12:30

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Published on: February 9, 2017

12.8K

Related Experiment Videos

Last Updated: Apr 16, 2026

Absolute Quantum Yield Measurement of Powder Samples
14:20

Absolute Quantum Yield Measurement of Powder Samples

Published on: May 12, 2012

28.9K
High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

11.5K
Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System
12:30

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Published on: February 9, 2017

12.8K

Area of Science:

  • Analytical Chemistry
  • Biophysics
  • Spectroscopy

Background:

  • Accurate determination of fluorescence quantum yields (QYs) is essential for photophysical characterization.
  • Existing methods can be time-consuming and require significant sample amounts.
  • Single-molecule spectroscopy offers high sensitivity for analyzing small sample volumes.

Purpose of the Study:

  • To develop a comparative method for accurate QY determination using fluorescence correlation spectroscopy (FCS).
  • To enable quantification of static and collisional quenching constants.
  • To facilitate photophysical characterization of labeled biomolecules under application-relevant conditions with low sample consumption.

Main Methods:

  • Utilizing fluorescence correlation spectroscopy (FCS) for sensitive QY determination.
  • Employing single-molecule spectroscopy techniques.
  • Combining FCS with fluorescence lifetime measurements.

Main Results:

  • Accurate determination of QYs for samples in the microliter range at (sub)nanomolar concentrations.
  • Quantification of both static and collisional quenching constants.
  • Demonstration of a simple, fast, and low-sample-consumption method.

Conclusions:

  • The presented FCS-based method provides accurate QY determination.
  • This technique is suitable for photophysically characterizing labeled biomolecules under demanding conditions.
  • The method's sensitivity and low sample requirement are advantageous for single-molecule studies.