Single Cell Metabolite Detection Using Inertial Microfluidics-Assisted Ion Mobility Mass Spectrometry

Area of Science:

• Analytical Chemistry
• Biotechnology
• Cell Biology

Background:

• Single-cell metabolite measurement is difficult due to isolation and detection challenges.
• Low metabolite levels hinder accurate identification and analysis.
• Existing methods lack efficiency and confidence in metabolite identification.

Purpose of the Study:

• To develop a novel strategy for single-cell metabolite detection and identification.
• To integrate spiral inertial microfluidics with ion mobility mass spectrometry (IM-MS).
• To enhance confidence in metabolite identification through gas-phase collisional cross section (CCS) measurements.

Main Methods:

• Cells in methanol suspension were focused using spiral inertial microfluidics.
• Focused cells were lysed and ionized via nanoelectrospray for real-time IM-MS analysis.
• Gas-phase collisional cross section (CCS) measurements were incorporated for enhanced identification.

Main Results:

• The system achieved six to eight single-cell metabolic fingerprints per minute.
• 19 distinct lipid species were identified in three cancer cell lines (U2OS, HepG2, HepG2.215) using CCS filtering.
• Principal component analysis (PCA) differentiated cancer cell lines based on surface phospholipids.

Conclusions:

• The developed technology platform provides an efficient method for single-cell lipid profiling.
• Ion mobility separation significantly improves the confidence of metabolite identification.
• This approach offers a promising tool for advancing single-cell metabolomics research.