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Real-Time, Two-Color Stimulated Raman Scattering Imaging of Mouse Brain for Tissue Diagnosis
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Real-Time Microscale Temperature Imaging by Stimulated Raman Scattering.

Benjamin Figueroa1, Ruoqian Hu1, Samuel G Rayner2,3

  • 1Department of Chemistry, University of Washington, Seattle, Washington 98195, United States.

The Journal of Physical Chemistry Letters
|August 14, 2020
PubMed
Summary

This study introduces a label-free stimulated Raman scattering (SRS) microscopy method for precise microscale temperature measurement in aqueous solutions. The technique accurately quantifies temperature changes in real-time without needing fluorescent probes.

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Area of Science:

  • Biophysics
  • Chemical Physics
  • Optical Microscopy

Background:

  • Accurate microscale thermometry is crucial for understanding heat dynamics in chemical and biological systems.
  • Existing fluorescent probes for temperature mapping have limitations due to microenvironment-dependent properties.

Purpose of the Study:

  • To develop a label-free ratiometric stimulated Raman scattering (SRS) microscopy technique for precise microscale temperature quantification.
  • To overcome the limitations of fluorescent probes in microscale thermometry.

Main Methods:

  • Utilized ratiometric stimulated Raman scattering (SRS) microscopy.
  • Monitored the O-H Raman stretching modes of water, specifically the ratio changes between the hydrogen-bonding O-H band and the isosbestic band.
  • Applied the technique for real-time, label-free temperature measurement.

Main Results:

  • Successfully quantified microscale temperature in real-time without exogenous contrast agents.
  • Demonstrated real-time measurement of localized intracellular and extracellular temperature changes induced by laser absorption.
  • Achieved submicrometer resolution for temperature mapping.

Conclusions:

  • The developed SRS microscopy technique provides accurate, label-free microscale thermometry for aqueous solutions.
  • This high-speed nonlinear optical imaging method offers potential for in situ imaging of thermogenesis in chemical and biological systems.