Abstract
Single-cell analysis of metabolites by mass spectrometry (MS) is challenging due to the very limited volume and inherent molecular complexity of the sample. Quantitative metabolomic analysis of individual cells provides information on the metabolic heterogeneity of cells unattainable by aggregate analysis of multiple cells. Depending on the ionization method, MS can offer quantitative analysis for a broad class of metabolites exhibiting both high sensitivity and selectivity. Laser ablation electrospray ionization (LAESI) has been successfully exploited to analyze metabolites from broad range of biological samples, including single cells and small cell populations. In this work, we describe a protocol for the relative quantitation of metabolites in single cells by LAESI-mass spectrometry.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Wang D, Bodovitz S (2010) Single cell analysis: the new frontier in “omics”. Trends Biotechnol 28:281–290
Lin Y, Trouillon R, Safina G, Ewing AG (2011) Chemical analysis of single cells. Anal Chem 83:4369–4392
Romanova EV, Rubakhin SS, Monroe EB, Sweedler JV (2009) Single cell mass spectrometry. In: Anselmetti D (ed.) Single cell analysis. Wiley-VCH, GmbH & Co. KGaA, Weinheim, pp 109–133
Li L, Golding RE, Whittal RM (1996) Analysis of single mammalian cell lysates by mass spectrometry. J Am Chem Soc 118:11662–11663
Jimenez CR, Veelen PAV, Li KW, Wildering WC, Geraerts WPM, Tjaden UR, Greef JVD (1994) Rapid communication: neuropeptide expression and processing as revealed by direct matrix-assisted laser desorption ionization mass spectrometry of single neurons. J Neurochem 62:404–407
Mizuno H, Tsuyama N, Harada T, Masujima T (2008) Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification. J Mass Spectrom 43:1692–1700
Lapainis T, Rubakhin SS, Sweedler JV (2009) Capillary electrophoresis with electrospray ionization mass spectrometric detection for single-cell metabolomics. Anal Chem 81: 5858–5864
Shrestha B, Vertes A (2009) In situ metabolic profiling of single cells by laser ablation electrospray ionization mass spectrometry. Anal Chem 81:8265–8271
Shrestha B, Nemes P, Vertes A (2010) Ablation and analysis of small cell populations and single cells by consecutive laser pulses. Appl Phys A Mater Sci Process 101:121–126
Shrestha B, Vertes A (2010) Direct analysis of single cells by mass spectrometry at atmospheric pressure. J Vis Exp 43:e2144
Sripadi P, Shrestha B, Easley RL, Carpio L, Kehn-Hall K, Chevalier S, Mahieux R, Kashanchi F, Vertes A (2010) Direct detection of diverse metabolic changes in virally transformed and tax-expressing cells by mass spectrometry. PLoS One 5:e12590
Parsiegla G, Shrestha B, Carrière F, Vertes A (2011) Direct analysis of phycobilisomal antenna proteins and metabolites in small cyanobacterial populations by laser ablation electrospray ionization mass spectrometry. Anal Chem 84:34–38
Shrestha B, Patt JM, Vertes A (2011) In situ cell-by-cell imaging and analysis of small cell populations by mass spectrometry. Anal Chem 83:2947–2955
Nemes P, Vertes A (2007) Laser ablation electrospray ionization for atmospheric pressure, in vivo, and imaging mass spectrometry. Anal Chem 79:8098–8106
Nemes P, Knolhoff AM, Rubakhin SS, Sweedler JV (2011) Metabolic differentiation of neuronal phenotypes by single-cell capillary electrophoresis–electrospray ionization-mass spectrometry. Anal Chem 83:6810–6817
Lu W, Bennett BD, Rabinowitz JD (2008) Analytical strategies for LC–MS-based targeted metabolomics. J Chromatogr B 871:236–242
Soga T, Ohashi Y, Ueno Y, Naraoka H, Tomita M, Nishioka T (2003) Quantitative metabolome analysis using capillary electrophoresis mass spectrometry. J Proteome Res 2:488–494
Kitteringham NR, Jenkins RE, Lane CS, Elliott VL, Park BK (2009) Multiple reaction monitoring for quantitative biomarker analysis in proteomics and metabolomics. J Chromatogr B 877:1229–1239
Ifa DR, Manicke NE, Rusine AL, Cooks RG (2008) Quantitative analysis of small molecules by desorption electrospray ionization mass spectrometry from polytetrafluoroethylene surfaces. Rapid Commun Mass Spectrom 22:503–510
Acknowledgments
The authors acknowledge financial support from the US National Science Foundation (Grant CHE-1152302) and the George Washington University Selective Excellence Fund for this work. The GeO2-based optical fibers were kindly donated by Infrared Fiber Systems for this study.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Shrestha, B., Vertes, A. (2014). Relative Quantitation in Single-Cell Metabolomics by Laser Ablation Electrospray Mass Spectrometry. In: Sriram, G. (eds) Plant Metabolism. Methods in Molecular Biology, vol 1083. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-661-0_3
Download citation
DOI: https://doi.org/10.1007/978-1-62703-661-0_3
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-660-3
Online ISBN: 978-1-62703-661-0
eBook Packages: Springer Protocols