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Related Concept Videos

Raman Spectroscopy: Overview01:20

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The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
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Raman Spectroscopy Instrumentation: Overview01:26

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A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
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Liquid-Liquid Phase Separation from the Viewpoint of Molecular Crowding Environment: A Raman Imaging Study.

Takakazu Nakabayashi1, Shinya Tahara1, Shinji Kajimoto1

  • 1Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan.

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Summary

This study quantifies biomolecular concentrations in cellular liquid droplets using Raman imaging. Droplet concentration varies with environment, impacting cellular molecular crowding and potential aggregation.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Chemical Physics

Background:

  • Molecular crowding environments are crucial for cellular processes.
  • Liquid-liquid phase separation (LLPS) forms intracellular droplets, influencing molecular crowding.
  • Quantifying biomolecular concentrations within these droplets is challenging.

Purpose of the Study:

  • To develop and apply an in situ method for quantifying biomolecular concentrations in cellular and in vitro liquid droplets.
  • To assess intracellular molecular crowding environments.
  • To investigate the relationship between droplet formation, biomolecular concentration, and aggregation.

Main Methods:

  • Raman imaging was employed to analyze molecular crowding.
  • Water's Raman band served as an internal intensity standard for quantification.
  • Biomolecular concentrations were measured in liquid droplets within buffer solutions and living cells.

Main Results:

  • In vitro liquid droplets exhibit high biomolecular concentrations (millimolar range).
  • Droplet concentration is sensitive to the surrounding environment, influencing droplet formation and potential aggregation.
  • In living cells, droplet biomolecular concentration is comparable to the surrounding cytoplasm, indicating selective molecular partitioning.

Conclusions:

  • Raman imaging provides a quantitative method to assess cellular molecular crowding.
  • LLPS in cells leads to biomolecular redistribution, creating concentrated environments within droplets.
  • Understanding these dynamics is key to comprehending cellular organization and function.