Single-Cell Lipidomics: An Automated and Accessible Microfluidic Workflow Validated by Capillary Sampling

Overview

Journal Anal Chem

Specialty Chemistry

Date 2024 Oct 26

PMID 39460701

Authors

Anastasia Kontiza

Johanna von Gerichten

Kyle D G Saunders

Matt Spick

Anthony D Whetton

Carla F Newman

Melanie J Bailey

Affiliations

Soon will be listed here.

Abstract

We report the first demonstration of a microfluidics-based approach to measure lipids in single living cells using widely available liquid chromatography mass spectrometry (LC-MS) instrumentation. The method enables the rapid sorting of live cells into liquid chambers formed on standard Petri dishes and their subsequent dispensing into vials for analysis using LC-MS. This approach facilitates automated sampling, data acquisition, and analysis and carries the additional advantage of chromatographic separation, aimed at reducing matrix effects present in shotgun lipidomics approaches. We demonstrate that our method detects comparable numbers of features at around 200 lipids in populations of single cells versus established live single-cell capillary sampling methods and with greater throughput, albeit with the loss of spatial resolution. We also show the importance of optimization steps in addressing challenges from lipid contamination, especially in blanks, and demonstrate a 75% increase in the number of lipids identified. This work opens up a novel, accessible, and high-throughput way to obtain single-cell lipid profiles and also serves as an important validation of single-cell lipidomics through the use of different sampling methods.

Citing Articles

Single-cell lipidomics: protocol development for reliable cellular profiling using capillary sampling.

Kontiza A, von Gerichten J, Spick M, Fraser E, Costa C, Saunders K Analyst. 2025; .

PMID: 40052368 PMC: 11886952. DOI: 10.1039/d5an00037h.

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