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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.
Super-resolution Fluorescence Microscopy01:37

Super-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.
UV–Vis Spectrometers01:14

UV–Vis Spectrometers

The absorbance of UV and visible (UV–visible) radiations is measured using a UV–visible spectrophotometer. Deuterium lamps, which emit UV radiation, and tungsten lamps, which produce radiation in the visible region, are used as light sources in UV–visible spectrophotometers. A monochromator or prism is used for diffraction grating, i.e., to split the incoming radiation into different wavelengths. A system of slits is used to focus the desired wavelength on the sample cell. Samples for...
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...
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,...
UV–Vis Spectroscopy: Molecular Electronic Transitions01:16

UV–Vis Spectroscopy: Molecular Electronic Transitions

In Ultraviolet–Visible (UV–Vis) spectroscopy, the absorption of electromagnetic radiation is used to probe the electronic structure of molecules. This technique provides insights into molecular electronic transitions, particularly the movement of electrons between different molecular orbitals. Radiation is absorbed if the energy of the electromagnetic radiation passing through the molecule is precisely equal to the energy difference between the excited and ground states. During this process,...

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Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
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Published on: August 29, 2025

A compact, multidimensional spectrofluorometer exploiting supercontinuum generation.

Hugh B Manning1, Gordon T Kennedy, Dylan M Owen

  • 1Chemical Biology Centre, Department of Chemistry, Imperial College London, London, UK. hugh.manning05@imperial.ac.uk

Journal of Biophotonics
|April 4, 2009
PubMed
Summary
This summary is machine-generated.

A new multidimensional spectrofluorometer offers advanced analysis of molecular interactions and tissue properties. This compact, automated instrument utilizes ultrafast supercontinuum light for detailed photophysical studies.

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Published on: December 1, 2023

Area of Science:

  • Biophysics
  • Spectroscopy
  • Optical Instrumentation

Background:

  • Characterizing photophysical properties is crucial for understanding molecular interactions and biological processes.
  • Existing spectrofluorometers may lack the resolution or versatility for comprehensive analysis.
  • Advanced instrumentation is needed for in situ measurements in diverse applications.

Purpose of the Study:

  • To report a novel, compact, and automated multidimensional spectrofluorometer.
  • To demonstrate its capability in studying molecular photophysics and interactions.
  • To showcase its application in characterizing complex biological samples like tissue autofluorescence.

Main Methods:

  • Development of a multidimensional spectrofluorometer.
  • Utilizing a fibre-laser-pumped ultrafast supercontinuum source.
  • Employing a fibre-optic probe for in situ measurements.

Main Results:

  • The instrument provides resolution in intensity, excitation/emission wavelength, decay time, and polarization.
  • Successfully studied the photophysics of the membrane probe di-4-ANEPPDHQ.
  • Characterized protein-protein interactions using Förster resonance energy transfer (FRET).
  • Demonstrated comprehensive characterization of tissue autofluorescence.

Conclusions:

  • The developed spectrofluorometer is a powerful tool for detailed photophysical and biochemical analysis.
  • Its compact and automated nature, coupled with in situ capabilities, broadens its applicability in medical and other fields.
  • This instrument enables a comprehensive understanding of complex biological systems through advanced spectroscopic measurements.