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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

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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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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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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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In chromatography, a solute moves through a chromatographic column and tends to spread, forming a Gaussian-shaped band. The longer the solute spends in the column, the broader the band becomes. The broadening can lead to overlaps within the column, affecting separation effectiveness.
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A racemic mixture, or racemate, is an equimolar mixture of enantiomers of a molecule that can be separated using their unique interaction with chiral molecules or media. Racemic mixtures are denoted by the (±)- prefix. This ‘optical rotation descriptor’ applies to the whole solution of a racemic mixture rather than a specific stereoisomer. Enantiomers typically have the same physical and chemical properties. Hence, they are not easily separable. However, enantiomers can exhibit...
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Super-Resolution Live Cell Imaging of Subcellular Structures
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Live-Cell Super-resolution Fluorescence Microscopy.

A S Mishin1, K A Lukyanov2

  • 1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia. mishin@ibch.ru.

Biochemistry. Biokhimiia
|June 20, 2019
PubMed
Summary
This summary is machine-generated.

Super-resolution fluorescence microscopy (nanoscopy) offers high-resolution imaging but struggles with live cells. This review covers nanoscopy methods and fluorescent labeling to minimize light damage for live-sample studies.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Optical microscopy is limited by diffraction.
  • Super-resolution microscopy (nanoscopy) surpasses this limit.
  • Current nanoscopy methods are challenging for live-cell imaging.

Purpose of the Study:

  • To review nanoscopy techniques.
  • To discuss fluorescent labeling strategies.
  • To address challenges in live-cell nanoscopy.

Main Methods:

  • Review of super-resolution microscopy techniques.
  • Analysis of fluorescent labeling for reduced phototoxicity.
  • Examination of methods for live biological samples.

Main Results:

  • Nanoscopy provides significantly enhanced spatial resolution.
  • Specific labeling reduces light-induced damage to cells.
  • Methods are being developed for live-cell applications.

Conclusions:

  • Advancements in nanoscopy and labeling are crucial for live-cell imaging.
  • Minimizing phototoxicity is key to studying dynamic cellular processes.
  • Further research is needed to optimize live-cell nanoscopy.