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Related Experiment Video

Updated: Oct 13, 2025

Applications of the Single-probe: Mass Spectrometry Imaging and Single Cell Analysis under Ambient Conditions
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High-Resolution Multi-Isotope Imaging Mass Spectrometry (MIMS) Imaging Applications in Stem Cell Biology.

G McMahon1, C P Lechene2

  • 1National Physical Laboratory, Teddington, United Kingdom.

Current Protocols
|November 17, 2021
PubMed
Summary
This summary is machine-generated.

Multi-isotope imaging mass spectrometry (MIMS) enables high-resolution analysis of molecular turnover in cells and tissues. This protocol details MIMS methods for stable isotope labeling and analysis, particularly for stem cell research.

Keywords:
BrdUMIMSNanoSIMSOpenMIMSstable isotopesthymidine

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Multi-isotope imaging mass spectrometry (MIMS) offers nanoscale spatial resolution for studying molecular turnover.
  • Stable isotopes like 15N, 13C, 18O, and 2H are crucial for tracing molecular dynamics.
  • Applications in stem cell research require specific labeling strategies.

Purpose of the Study:

  • To provide a practical protocol for MIMS analysis.
  • To detail stable isotope labeling techniques for DNA in cells and animals.
  • To guide sample preparation, high-resolution imaging, and data analysis for MIMS.

Main Methods:

  • Stable isotope labeling of DNA using 15N-thymidine, 13C-thymidine, or BrdU (81Br).
  • Sample preparation for secondary ion mass spectrometry (SIMS) using Cameca NanoSIMS 50L.
  • High-resolution imaging and statistical analysis of label incorporation in regions of interest.

Main Results:

  • Demonstration of MIMS for analyzing DNA replication and turnover in single cells.
  • Methodology for robust quantification of stable isotope incorporation.
  • Successful application of MIMS in stem cell research contexts.

Conclusions:

  • MIMS is a powerful technique for quantitative molecular analysis at the nanoscale.
  • The protocol provides a comprehensive guide for implementing MIMS in biological research.
  • This method facilitates detailed studies of cellular processes, especially in stem cells.