Global and Spatial Metabolomics of Individual Cells Using a Tapered Pneumatically Assisted nano-DESI Probe

Area of Science:

• Analytical Chemistry
• Cell Biology
• Metabolomics

Background:

• Single-cell metabolomics offers deep insights into cellular mechanisms but faces challenges in metabolite detection due to chemical properties and concentration variability.
• Existing methods struggle with the sensitivity and specificity required for analyzing metabolites within individual cells.

Purpose of the Study:

• To develop and validate a novel analytical approach for high-throughput single-cell metabolomics.
• To enable detailed chemical imaging and global metabolomics of both large and small individual cells.
• To investigate cellular heterogeneity and metabolic responses at the single-cell level.

Main Methods:

• Development of a tapered probe for pneumatically assisted nanospray desorption electrospray ionization (PA nano-DESI) mass spectrometry.
• Application of the PA nano-DESI probe for chemical imaging of larger cells and global metabolomics of 15 μm cells.
• Establishment of predefined cell arrays for rapid, high-throughput analysis of individual cells (3 cells/min).

Main Results:

• Successful acquisition of metabolomic data from 479 individual cells using the PA nano-DESI probe.
• Optimization of analytical conditions demonstrated through analysis of multiple cells.
• Monitoring of metabolome alterations in INS-1 cells exposed to varying glucose concentrations (93-97 cells analyzed per condition).

Conclusions:

• The developed PA nano-DESI probe significantly advances single-cell metabolomics capabilities.
• This approach provides valuable insights into cellular heterogeneity and dynamic metabolic processes.
• The method enables comprehensive analysis of cellular responses to stimuli at an unprecedented scale.