Single-Cell Untargeted Lipidomics Using Liquid Chromatography and Data-Dependent Acquisition after Live Cell...

Area of Science:

• Analytical Chemistry
• Cell Biology
• Biochemistry

Background:

• Lipids are crucial for cellular structure, energy, and signaling.
• Understanding single-cell lipid heterogeneity is vital for biological research.
• Existing methods lack the sensitivity for single-cell lipid analysis.

Purpose of the Study:

• To develop and validate an untargeted single-cell lipidomics method.
• To demonstrate the method's sensitivity for lipid identification and cell classification.
• To analyze dynamic changes in single-cell lipid profiles.

Main Methods:

• Microflow chromatography coupled to data-dependent mass spectrometry.
• Method validation using dilute cell extracts and standards.
• Automated live single-cell sampling with spatial and morphological retention.

Main Results:

• Successful development and validation of a sensitive single-cell lipidomics workflow.
• Demonstrated lipid profile heterogeneity across single cells.
• Observed changes in single-cell lipid profiles and lipid droplet expansion in response to oxidative stress.

Conclusions:

• The developed method enables sensitive, untargeted lipidomics at the single-cell level.
• The workflow can capture dynamic lipid changes and cellular responses to stimuli.
• This advancement will facilitate deeper understanding of lipid roles in diverse biological processes.