Identification of Metabolites in Single Cells by Ion Mobility Separation and Mass Spectrometry

Linwen Zhang; Linda L Allworth; Akos Vertes

doi:10.1007/978-1-4939-9831-9_2

Identification of Metabolites in Single Cells by Ion Mobility Separation and Mass Spectrometry

Methods Mol Biol. 2020:2064:9-18. doi: 10.1007/978-1-4939-9831-9_2.

Authors

Linwen Zhang^{1

2}, Linda L Allworth³, Akos Vertes⁴

Affiliations

¹ Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MB, USA.
² Institute for Bioscience and Biotechnology Research, Rockville, MB, USA.
³ Thomas Jefferson High School for Science and Technology, Alexandria, VA, USA.
⁴ Department of Chemistry, The George Washington University, Washington, DC, USA. vertes@gwu.edu.

PMID: 31565763
DOI: 10.1007/978-1-4939-9831-9_2

Abstract

Non-targeted metabolic analysis of single cells by mass spectrometry (MS) is important for understanding individual cell functions and characterizing cell-to-cell heterogeneity. However, identifying biomolecules in single cells presents significant challenges due to the low picoliter volume samples and the structural diversity of metabolites. Capillary microsampling electrospray ionization (ESI) MS with ion mobility separation (IMS) enables the analysis of single cells under ambient conditions with minimum sample pretreatment and improved specificity. Here, we describe a protocol for the analysis of the metabolic makeup, and the identification of ions produced from single cells by capillary microsampling ESI-IMS-MS.

Keywords: Collision cross-section; Ion mobility separation; Isobaric ions; Mass spectrometry; Metabolomics; Single-cell analysis.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Hep G2 Cells
Humans
Metabolome
Metabolomics / methods*
Single-Cell Analysis / methods*
Spectrometry, Mass, Electrospray Ionization / methods*