Frequency Modulation-Boosted Electrolaunching Ionization Mass Spectrometry Enables Mass-Selective Single-Cell...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Cell Biology

Background:

• Bulk cell analysis limits understanding of cellular heterogeneity in disease.
• Intact living-cell electrolaunching ionization mass spectrometry (ILCEI-MS) improves sensitivity but has limitations.
• Single-cell chemical profiling offers deeper insights into molecular mechanisms of disease.

Purpose of the Study:

• To develop a mass-selective single-cell metabolomics approach using induced ILCEI-MS.
• To enhance ionization efficiency, matrix tolerance, and enable mass-selective detection.
• To demonstrate the platform's capability for analyzing cellular heterogeneity.

Main Methods:

• Developed induced ILCEI-MS with frequency-modulated AC voltages for mass-selective detection.
• Eliminated physical contact between cells and electrodes to prevent interference.
• Analyzed single intact GL261 cells, achieving high throughput and extensive ion coverage.

Main Results:

• Achieved high single-cell detection throughput (∼45 cells/min) and broad ion coverage (∼400 ions/cell).
• Demonstrated AC frequency-dependent mass-selective detection: lower frequencies for higher m/z ions, higher frequencies for lower m/z ions.
• Successfully differentiated various cell types and subtypes, showcasing potential for heterogeneity studies.

Conclusions:

• The developed method provides a powerful tool for mass-selective single-cell metabolomics.
• Frequency-modulated AC voltage enables targeted ion analysis for comprehensive single-cell profiling.
• This approach significantly advances the study of cellular heterogeneity and disease pathogenesis.