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Three-dimensional electron paramagnetic resonance imaging of mice using ascorbic acid sensitive nitroxide imaging

Hideo Sato-Akaba1, Miho C Emoto2, Ken-Ichi Yamada3

  • 1Department of Systems Innovation, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka, Japan.

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Summary

Researchers developed an advanced electron paramagnetic resonance (EPR) imager to overcome limitations in visualizing fast-reacting nitroxide probes for in vivo redox imaging. This breakthrough enables sensitive assessment of oxidative stress using previously unsuitable probes.

Keywords:
EPRRedox statusblood brain barrierimagingnitroxidereactive oxygen species (ROS)

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Area of Science:

  • Biomedical Imaging
  • Chemical Biology
  • Biophysics

Background:

  • Nitroxide compounds are redox-sensitive probes for in vivo oxidative stress assessment using electron paramagnetic resonance (EPR).
  • Rapid reduction of some nitroxides in vivo limits their use with existing EPR imagers due to slow operational speeds.
  • Developing faster EPR imaging is crucial for utilizing a wider range of nitroxide probes.

Purpose of the Study:

  • To enhance EPR system speed and sensitivity for improved 3D imaging of nitroxide probes in vivo.
  • To assess the capability of the improved EPR imager using nitroxide probes with varying reduction rates.
  • To demonstrate the potential for visualizing dynamic biological processes with high redox sensitivity.

Main Methods:

  • An advanced EPR system was developed with improved stability and sensitivity for 3D imaging.
  • Two nitroxide probes, 4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (Tempol) and 2,6-dispiro-4",4"-dipyrane-piperidine-4-one-N-oxyl (DiPy), were used.
  • 3D EPR imaging was performed on mouse brains and heads following probe administration.

Main Results:

  • The improved EPR imager successfully acquired clear 3D EPR images of mouse brains and heads.
  • Tempol was visualized, confirming its presence with an unpaired electron in vivo.
  • The imager captured the dynamic process of DiPy entering, distributing, and being reduced within the mouse brain within seconds.

Conclusions:

  • The enhanced EPR imager overcomes limitations of probe reduction speed, enabling sensitive in vivo redox assessment.
  • This technology allows for the use of previously unsuitable nitroxide probes for imaging.
  • The improved system opens new possibilities for studying dynamic redox processes in biological systems.