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Related Concept Videos

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.
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,...
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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.

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Updated: Jun 17, 2026

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals
07:34

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals

Published on: August 22, 2019

Full spectrum filterless fluorescence microscopy.

M J Booth1, A Jesacher, R Juskaitis

  • 1Department of Engineering Science, University of Oxford Parks Road, Oxford OX13PJ, United Kingdom. martin.booth@eng.ox.ac.uk

Journal of Microscopy
|January 9, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces a novel fluorescence microscope capable of capturing full spectrum images across a wide wavelength range. This versatile system overcomes limitations of conventional microscopes, enabling simultaneous excitation and emission spectrum detection for enhanced biological imaging.

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Area of Science:

  • Microscopy
  • Spectroscopy
  • Biophotonics

Background:

  • Conventional fluorescence microscopy separates excitation and emission using the Stokes shift and filters.
  • This spectral separation limits the ability to capture the full excitation/emission spectrum in a single image.
  • Existing methods restrict comprehensive spectral analysis of fluorescent specimens.

Purpose of the Study:

  • To develop a versatile fluorescence microscope system.
  • To enable simultaneous acquisition of full excitation and emission spectra.
  • To improve background rejection and enable 3D imaging.

Main Methods:

  • Developed a fluorescence microscope with illumination across 450-680 nm.
  • Enabled simultaneous detection of the full emission spectrum within the same range.
  • Integrated structured illumination optical sectioning for 3D imaging and background rejection.

Main Results:

  • Demonstrated full spectrum fluorescence imaging of biological specimens.
  • Achieved simultaneous detection of excitation and emission spectra.
  • Obtained 3D resolved images with enhanced background rejection.

Conclusions:

  • The novel microscope offers greater versatility than standard fluorescence microscopes.
  • This system has broad applicability in various fluorescence imaging applications.
  • Enables comprehensive spectral analysis previously not possible in a single image.