Lipidomic profiling of single mammalian cells by infrared matrix-assisted laser desorption electrospray ionization (IR-M
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RESEARCH PAPER
Lipidomic profiling of single mammalian cells by infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) Ying Xi 1 & Anqi Tu 1 & David C. Muddiman 1,2 Received: 10 July 2020 / Revised: 11 September 2020 / Accepted: 18 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract To better understand cell-to-cell heterogeneity, advanced analytical tools are in a growing demand for elucidating chemical compositions of each cell within a population. However, the progress of single-cell chemical analysis has been restrained by the limitations of small cell volumes and minute cellular concentrations. Here, we present a rapid and sensitive method for investigating the lipid profiles of isolated single cells using infrared matrix-assisted laser desorption electrospray ionization mass spectrometry (IR-MALDESI-MS). In this work, HeLa cells were dispersed onto a glass slide, and the cellular contents were ionized by IR-MALDESI and measured using a Q-Exactive HF-X mass spectrometer. Importantly, this approach does not require extraction and/or enrichment of analytes prior to MS analysis. Using this approach, 45 distinct lipid species, predominantly phospholipids, were detected and putatively annotated from the single HeLa cells. The proof-of-concept study demonstrates the feasibility and efficacy of IR-MALDESI-MS for rapid lipidomic profiling of single cells, which provides an important basis for future work on differentiation between normal and diseased cells at various developmental states, which can offer new insights into cellular metabolic pathways and pathological processes. Although not yet accomplished, we believe this approach can be readily used as an assessment tool to compare the number of identified species during source evolution and method optimization (intra-laboratory), and also disclose the complementary nature of different direct analytical approaches for the coverage of different types of endogenous analytes (inter-laboratory). Keywords Single-cell analysis . Lipidome . Mass spectrometry . IR-MALDESI . Orbitrap
Introduction Heterogeneity is an intrinsic attribute, existing everywhere in biological systems. It is reported that even cells originating from the same genome have diverse chemical and physiological features because of their different microenvironments and Ying Xi and Anqi Tu contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02961-6) contains supplementary material, which is available to authorized users. * David C. Muddiman [email protected] 1
FTMS Laboratory for Human Health Research, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA
2
Molecular Education, Technology and Research Innovation Center (METRIC), North Carolina State University, Raleigh, NC 27695, USA
stochastic processes [1–3]. These unicellular properties carry critical biological information regarding cell activities and molecular functions [4], f
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