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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.
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.
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,...

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Related Experiment Video

Updated: Jul 10, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Beyond the diffraction limit: far-field fluorescence imaging with ultrahigh resolution.

James H Rice1

  • 1School of Chemical Sciences and Pharmacy, University of East Anglia, Earlham Road, Norwich, UK NR2 3RG. james.rice@uea.ac.uk

Molecular Biosystems
|October 18, 2007
PubMed
Summary

Researchers are overcoming the diffraction limit in fluorescence microscopy. New far-field techniques now enable ultrahigh resolution imaging for biological applications, pushing beyond traditional limits.

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Last Updated: Jul 10, 2026

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Area of Science:

  • Biophysics
  • Optical Imaging
  • Cell Biology

Background:

  • Fluorescence microscopy is crucial for biological imaging but limited by diffraction.
  • Overcoming the diffraction limit is a key challenge in microscopy.

Purpose of the Study:

  • To review advancements in far-field fluorescence microscopy for ultrahigh resolution.
  • To discuss the biological applications of these advanced imaging techniques.

Main Methods:

  • Focuses on far-field fluorescence microscopy techniques.
  • Highlights methods that surpass the diffraction limit.

Main Results:

  • Emerging methods achieve fluorescence imaging resolution well beyond the diffraction limit.
  • These techniques are being applied to various biological systems.

Conclusions:

  • Far-field fluorescence imaging offers unprecedented resolution for biological studies.
  • Future trends point towards continued innovation in ultrahigh-resolution microscopy.