Metabolic Analysis at the Nanoscale with Multi-Isotope Imaging Mass Spectrometry (MIMS)

Area of Science:

• Cell Biology
• Biophysics
• Biochemistry

Background:

• Stable-isotope tracers are crucial for understanding cellular metabolism.
• Existing imaging techniques have limitations in resolution and quantitative analysis of metabolic processes.
• Multi-isotope imaging mass spectrometry (MIMS) offers a powerful solution for high-resolution metabolic analysis.

Purpose of the Study:

• To introduce and demonstrate the application of metabolic analysis using NanoSIMS, a type of MIMS.
• To showcase the versatility of MIMS in biological research.
• To provide detailed protocols for metabolic MIMS experiments.

Main Methods:

• Metabolic labeling of cells or animals with stable-isotope tracers.
• Utilizing NanoSIMS for high-resolution isotope ratio measurements at the submicron level.
• Correlative microscopy techniques (TEM, light microscopy) for integrated analysis.

Main Results:

• Demonstrated quantification of protein turnover in single organelles.
• Successfully tracked cell replication in mouse tissues in vivo.
• Achieved submicron resolution for metabolic tracer localization and quantification.

Conclusions:

• Metabolic analysis with MIMS, particularly using NanoSIMS, is a versatile technique for biological research.
• MIMS provides unprecedented insights into cellular and tissue-level metabolic dynamics.
• The presented protocols facilitate the application of MIMS in diverse biological studies.