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bioRxiv · 10.1101/2022.02.15.479799

High-throughput detection and quantification of phosphatidylcholines and sphingomyelins from single cells by chip-based nanoelectrospray ionisation

Hancock, S. E.·Ding, E.·Johansson Beves, E.·Mitchell, T. W.·Turner, N.
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Abstract

Recent advances in single-cell genomics and transcriptomics technologies have transformed our understanding of cellular heterogeneity in growth, development, ageing and disease; however, methods for single-cell lipidomics have comparatively lagged behind in development. We have developed a high-throughput method for the detection and quantification of a wide range of phosphatidylcholine (PC) and sphingomyelin (SM) species from single cells that combines fluorescence-assisted cell sorting (FACS) with automated chip-based nanoelectrospray ionization (nanoESI) and shotgun lipidomics. We show herein that our method is capable of quantifying more than 50 different PC and SM species from single cells and can easily distinguish between cells of different lineages or cells treated with exogenous fatty acids. Moreover, our method can detect more subtle differences in the lipidome between cell lines of the same cancer type. Our approach can be run in parallel with other single-cell technologies to deliver near-complete multi-omics data on cells with a similar phenotype and has the capacity to significantly advance our current knowledge on cellular heterogeneity.

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Hancock, S. E., Ding, E., Johansson Beves, E., Mitchell, T. W., Turner, N.. 2022-02-16. High-throughput detection and quantification of phosphatidylcholines and sphingomyelins from single cells by chip-based nanoelectrospray ionisation. https://doi.org/10.1101/2022.02.15.479799

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