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¹³C NMR: ¹H–¹³C Decoupling01:04

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The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
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Real-Time Metabolic Detection in Living Cells Using Hyperpolarized 13C NMR
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Real-Time Metabolic Detection in Living Cells Using Hyperpolarized 13C NMR

Published on: July 8, 2025

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Real-Time Metabolic Detection in Living Cells Using Hyperpolarized 13C NMR.

Tomoto Ura1, Keita Saito2, Ryoma Kobayashi2

  • 1Institute for Quantum Life Science, National Institutes for Quantum Science and Technology; Institute of Pure and Applied Sciences, University of Tsukuba.

Journal of Visualized Experiments : Jove
|July 28, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a novel bioreactor system integrated with hyperpolarized 13C nuclear magnetic resonance (NMR) spectroscopy for real-time cellular metabolism analysis. The method enhances the study of pyruvate metabolism in live cells with improved accuracy and reproducibility.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Spectroscopy

Background:

  • Real-time analysis of cellular metabolism is crucial for understanding biological processes and disease states.
  • Conventional methods for measuring metabolic flux often lack temporal resolution or require cell lysis.
  • Hyperpolarized 13C nuclear magnetic resonance (NMR) spectroscopy offers a non-invasive approach to track metabolic pathways.

Purpose of the Study:

  • To develop and validate a method for real-time measurement of pyruvate metabolism in live cells using a custom-built bioreactor system coupled with hyperpolarized 13C NMR.
  • To establish a stable cell culture environment within the bioreactor for precise substrate injection and signal acquisition.
  • To assess the reproducibility and kinetic accuracy of the developed method for metabolic studies.

Main Methods:

  • Integration of an in-house-built bioreactor with a hyperpolarized 13C NMR spectroscopy setup.
  • Development of a protocol for precise timing of hyperpolarized substrate (pyruvate) injection and NMR signal acquisition.
  • Encapsulation of live cells within alginate gels for stable culture and placement within the bioreactor.
  • In situ mixing capability within the bioreactor to accelerate signal detection and minimize relaxation losses.

Main Results:

  • Reliable detection of pyruvate-to-lactate conversion within a 60-second window with a sufficient signal-to-noise ratio.
  • Demonstration of improved reproducibility and kinetic accuracy compared to conventional metabolic measurement techniques.
  • Successful maintenance of a stable cell culture environment throughout the real-time measurement process.

Conclusions:

  • The developed bioreactor-NMR system enables real-time, in vivo monitoring of cellular pyruvate metabolism with enhanced accuracy.
  • This method holds significant potential for applications in cancer metabolism research and high-throughput drug screening.
  • Future refinements, including exploring alternative gel matrices, could further expand the system's versatility for diverse biological studies.