Fluorescence Spectroscopy

Fluorescence spectroscopy is an analytical method that measures light emitted by a substance after it absorbs light, providing sensitive information about molecular composition and behavior. In this process, excitation light raises fluorescent molecules to a higher electronic state, and they release part of that energy as longer-wavelength emission when returning to a lower state; emission intensity and wavelength reflect the molecule’s environment and concentration. In biology, the technique supports protein and nucleic acid analysis, cellular imaging, and studies of molecular interactions. Fluorescent labels and reporter molecules also enable researchers to track biochemical processes in real time, advancing diagnostics, drug development, and cell biology.

Fluorescence Spectroscopy - Related Videos

Education

JoVE Core - Analytical Chemistry

Atomic Fluorescence Spectroscopy

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2024

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...

Research

JoVE Journal - Engineering

High Resolution Phonon-assisted Quasi-resonance Fluorescence Spectroscopy

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Cited by 2 •

2016

The manuscript describes a method of phonon-assisted quasi-resonant fluorescence spectroscopy that incorporates both laser-limited resolution and photoluminescence (PL) spectroscopy. This method utilizes optical phonons to provide linewidth-limited resolution spectra of atom-like semiconductor structures in the energy domain. The method is also easily realized with a single spectrometer optical spectroscopy setup.

Fluorescence Fluctuation Spectroscopy to Study Protein Homo-Oligomerization

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2025

This video demonstrates the technique of fluorescence fluctuation spectroscopy (FFS) to study protein homo-oligomerization. Fluorescently-labeled proteins in a sample are dimerized using an agent when studying protein oligomerization using FFS. Using a confocal microscope, as the proteins move in and out of the small observation volume, the analysis of the fluctuations in the brightness of the fluorescent molecules is performed to determine the oligomeric state of the proteins.

Fluorescence Fluctuation Spectroscopy to Study Protein Interaction at Cell Contacts

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2025

This video demonstrates the use of fluorescence fluctuation spectroscopy to detect the interaction among cell surface proteins at cell-cell contacts. By expressing the transmembrane adhesion receptor of interest labeled with a fluorescent protein and mixing two different cell populations harboring two spectrally separated fluorescent labels, the trans-interaction between the receptors of two neighboring cells with different-colored fluorescence is assessed via cross-correlation in the...

Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)

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Cited by 9 •

2012

A technique to probe the lipid raft partitioning of fluorescent proteins at the plasma membrane of living cells is described. It takes advantage of the disparity in diffusion times of proteins located inside or outside of lipid rafts. Acquisition can be performed dynamically in control conditions or after drug addition.

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