Single Cell Untargeted Lipidomics Using Liquid Chromatography Ion Mobility-Mass Spectrometry

Area of Science:

• Biochemistry
• Analytical Chemistry
• Cell Biology

Background:

• Omics analysis now achieves single cell resolution, with transcriptomics and genomics leading advancements.
• Single cell proteomics has rapidly progressed, but single cell lipidomics methods are scarce, especially non-imaging approaches.
• Single cell lipidomics offers potential for understanding cellular heterogeneity in diseases and enabling multiomics studies.

Purpose of the Study:

• To develop and demonstrate an untargeted single cell lipidomics methodology.
• To enhance lipid coverage and enable lipid profiling at the single cell level.
• To lay the groundwork for integrated single cell multiomics.

Main Methods:

• Development of untargeted single cell lipidomics using nanoflow liquid chromatography-ion mobility spectrometry-mass spectrometry (nLC-IMS-MS).
• Application of both positive and negative ion modes to maximize lipid identification.
• Analysis of human cholangiocarcinoma cells at the single cell level.

Main Results:

• Identification of an average of 161 lipids (phospholipids, sphingolipids, cholesteryl esters, glycerides) in positive ion mode.
• Identification of an average of 20 phospholipid species in negative ion mode.
• Demonstration of a novel method for lipid profiling from single or low cell inputs.

Conclusions:

• A new untargeted single cell lipidomics method has been successfully developed and validated.
• The methodology significantly enhances lipid coverage and enables detailed lipid analysis in single cells.
• This approach is crucial for advancing single cell multiomics and understanding cellular lipid heterogeneity.