Related Experiment Video
Updated: Jun 6, 2026
07:34
Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals
Published on: August 22, 2019
Encapsulated energy-transfer cassettes with extremely well resolved fluorescent outputs
Yuichiro Ueno1, Jiney Jose, Aurore Loudet
1Department of Chemistry, Texas A & M University, Box 30012, College Station, Texas 77841, USA.
Journal of the American Chemical Society
|November 26, 2010
Summary
Researchers developed new water-compatible fluorescent imaging probes using BODIPY and cyanine dyes. Encapsulated in nanoparticles, these probes enable efficient cellular imaging with tunable, single-wavelength excitation and red emission, localizing in mitochondria.
Area of Science:
- Chemical Biology
- Materials Science
- Biophysics
Background:
- Development of fluorescent imaging probes is crucial for biological research.
- Existing probes often lack tunable photonic properties or require multiple excitation wavelengths.
- Need for water-compatible probes with large spectral windows for multiplexing.
Purpose of the Study:
- To develop water-compatible fluorescent imaging probes with tunable photonic properties.
- To investigate energy transfer mechanisms in bichromophoric cassettes.
- To enable cellular imaging and organelle labeling using a single excitation source.
Main Methods:
- Synthesis of bichromophoric cassettes (BODIPY donor, cyanine acceptor).
- Photophysical characterization including excitation, emission, and energy transfer rate measurements.
- Encapsulation of probes into calcium phosphate/silicate nanoparticles.
- Cellular uptake and localization studies in rat liver cells.
Main Results:
- Bichromophoric cassettes exhibited efficient through-bond energy transfer, enabling large quasi-Stokes shifts (86–290 nm).
- Probes showed tunable emission at ~600, 700, and 800 nm upon single-wavelength excitation.
- Nanoparticle encapsulation improved water dispersibility with minimal impact on photophysical properties.
- Encapsulated probes successfully entered cells and localized in mitochondria, emitting red fluorescence.
Conclusions:
- Readily accessible cyanine-based cassettes facilitate efficient through-bond energy transfer for advanced imaging.
- Nanoparticle formulation provides water compatibility for biological applications.
- These probes are suitable for multiplexing and organelle-specific labeling in live cells.
Related Concept Videos
01:37Super-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.
01:25Fluorescence 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.