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Related Concept Videos

Atomic Fluorescence Spectroscopy01:29

Atomic Fluorescence Spectroscopy

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Atomic fluorescence spectroscopy (AFS) is an analytical technique that involves the electronic transitions of atoms in a flame, furnace, or plasma being excited by electromagnetic (EM) radiation. When these atoms absorb energy, they become excited and subsequently release energy as they return to their original state. This emitted light, or "fluorescence," is observed at a right angle to the incident beam. Both absorption and emission processes transpire at distinct wavelengths, which...
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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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Interferometric fluorescence cross correlation spectroscopy.

Ipsita Saha1,2, Saveez Saffarian1,2,3

  • 1Center for Cell and Genome Science, University of Utah, Salt Lake City, Utah, United States of America.

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|December 19, 2019
PubMed
Summary

We developed a new method using single photon interference and fluorescence correlation spectroscopy (FCS) to measure molecular transport in cells and materials. This technique rapidly quantifies diffusion and flow, overcoming common artifacts.

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Area of Science:

  • Biophysics
  • Materials Science
  • Chemical Physics

Background:

  • Understanding molecular transport is crucial for cellular dynamics and novel materials.
  • Fluorescent molecule concentration fluctuations reveal transport properties in heterogeneous systems.
  • Existing methods face challenges with speed, resolution, and artifacts like blinking.

Purpose of the Study:

  • To present a novel method for simultaneously measuring diffusion and directional flow of fluorescent molecules.
  • To enable rapid, high-resolution transport measurements in complex environments.
  • To address and mitigate photo-physical artifacts in fluorescence measurements.

Main Methods:

  • Utilizing single photon interference combined with fluorescence correlation spectroscopy (FCS).
  • Analyzing concentration fluctuations of fluorescent molecules in aqueous samples.
  • Measuring transport in thousands of 100 nm voxels within seconds.

Main Results:

  • Simultaneous measurement of diffusion and directional flow achieved.
  • Method validated using quantum dots and VSV-G receptor transport in cellular membranes and gels.
  • Elimination of photo-physical artifacts like blinking under specific conditions.

Conclusions:

  • The presented method offers a rapid and robust approach for quantifying molecular transport.
  • This technique is applicable to diverse systems, from biological membranes to synthetic materials.
  • The method enhances the study of dynamics in heterogeneous environments.