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Related Concept Videos

Photoluminescence: Applications01:14

Photoluminescence: Applications

Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
Transmission Electron Microscopy01:15

Transmission Electron Microscopy

In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400 keV in...
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.
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
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Overview of Electron Microscopy01:25

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
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.

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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Published on: August 7, 2016

Photooxidation technology for correlated light and electron microscopy.

C Meisslitzer-Ruppitsch1, C Röhrl, J Neumüller

  • 1Department of Cell Biology and Ultrastructure Research, Centre for Anatomy and Cell Biology, Medical University Vienna, Vienna, Austria.

Journal of Microscopy
|September 17, 2009
PubMed
Summary

Correlative microscopy bridges light and electron imaging. Photooxidation methods utilize fluorescent dyes to visualize cellular structures with high resolution, enhancing cell biology research.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biochemistry

Background:

  • Classical light and electron microscopy have distinct resolution limits.
  • Technical advancements are narrowing the gap between light and electron microscopy.
  • Correlative microscopy techniques offer synergistic advantages for biological imaging.

Purpose of the Study:

  • To highlight the utility of photooxidation methods in correlative microscopy.
  • To detail the application of fluorescent dyes and diaminobenzidine (DAB) for dual-level imaging.
  • To discuss methodical solutions for optimizing photooxidation-based fine structural localization.

Main Methods:

  • Utilizing fluorescent dyes to photooxidize diaminobenzidine (DAB).
  • Excitation of free oxygen radicals upon fluorochrome illumination initiates the reaction.
  • Osmium staining of DAB precipitates for electron microscopy visualization.

Main Results:

  • Photooxidation generates DAB precipitates, visible via electron microscopy.
  • These precipitates label cellular structures identified by fluorescent probes at the light microscopy level.
  • Successful fine structural localization depends on fluorochrome choice and reaction conditions.

Conclusions:

  • Photooxidation methods are valuable tools for correlative light and electron microscopy.
  • This technique enables high-resolution visualization of cellular structures.
  • Optimizing fluorochrome selection and reaction parameters is crucial for effective application.