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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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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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Super-resolution Fluorescence Microscopy01:37

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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...
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Total Internal Reflection Fluorescence Microscopy01:05

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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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Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Fluorescence and Phosphorescence: Instrumentation01:25

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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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Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Fluorescence (Multiwave) Confocal Microscopy.

J Welzel1, Raphaela Kästle1, Elke C Sattler2

  • 1Department of Dermatology and Allergology, General Hospital Augsburg, Sauerbruchstrasse 6, 86179 Augsburg, Germany.

Dermatologic Clinics
|October 4, 2016
PubMed
Summary
This summary is machine-generated.

Multiwave confocal microscopy uses different laser wavelengths and fluorophores for in vivo and ex vivo imaging. This technique enhances contrast for visualizing epithelial structures like tumors, nerves, and glands.

Keywords:
Fluorescence confocal microscopyFluorescent dyesFluorophoresPhotobleachingQuenchingSelective featuring of target structuresStaining

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Area of Science:

  • Biomedical Imaging
  • Optical Microscopy

Background:

  • Reflectance confocal microscopy is a standard imaging technique.
  • Limitations exist in contrast and specific structure visualization.

Purpose of the Study:

  • To introduce and evaluate fluorescence mode confocal microscopy using multiwave systems.
  • To demonstrate enhanced visualization of specific biological structures.

Main Methods:

  • Utilized multiwave confocal microscopes with varied laser wavelengths.
  • Employed exogenous fluorophores for fluorescence enhancement.
  • Performed in vivo and ex vivo imaging experiments.

Main Results:

  • Fluorescence mode confocal microscopy significantly improved contrast between epithelium and surrounding tissues.
  • Enabled clear depiction of epithelial tumors, nerves, and glands.
  • Demonstrated utility for both in vivo and ex vivo applications.

Conclusions:

  • Multiwave confocal microscopy with fluorescence offers superior contrast and structural detail.
  • This advanced technique provides valuable insights for diagnosing and studying epithelial tissues.