Skip to main content

Relative Quantitation in Single-Cell Metabolomics by Laser Ablation Electrospray Mass Spectrometry

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1083))

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.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Wang D, Bodovitz S (2010) Single cell analysis: the new frontier in “omics”. Trends Biotechnol 28:281–290

    Article  PubMed  CAS  Google Scholar 

  2. Lin Y, Trouillon R, Safina G, Ewing AG (2011) Chemical analysis of single cells. Anal Chem 83:4369–4392

    Article  PubMed  CAS  Google Scholar 

  3. 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

    Google Scholar 

  4. Li L, Golding RE, Whittal RM (1996) Analysis of single mammalian cell lysates by mass spectrometry. J Am Chem Soc 118:11662–11663

    Article  CAS  Google Scholar 

  5. 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

    Article  PubMed  CAS  Google Scholar 

  6. 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

    Article  PubMed  CAS  Google Scholar 

  7. Lapainis T, Rubakhin SS, Sweedler JV (2009) Capillary electrophoresis with electrospray ionization mass spectrometric detection for single-cell metabolomics. Anal Chem 81: 5858–5864

    Article  PubMed  CAS  Google Scholar 

  8. Shrestha B, Vertes A (2009) In situ metabolic profiling of single cells by laser ablation electrospray ionization mass spectrometry. Anal Chem 81:8265–8271

    Article  PubMed  CAS  Google Scholar 

  9. 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

    Article  CAS  Google Scholar 

  10. Shrestha B, Vertes A (2010) Direct analysis of single cells by mass spectrometry at atmospheric pressure. J Vis Exp 43:e2144

    Google Scholar 

  11. 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

    Article  PubMed  Google Scholar 

  12. 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

    Article  PubMed  Google Scholar 

  13. 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

    Article  PubMed  CAS  Google Scholar 

  14. Nemes P, Vertes A (2007) Laser ablation electrospray ionization for atmospheric pressure, in vivo, and imaging mass spectrometry. Anal Chem 79:8098–8106

    Article  PubMed  CAS  Google Scholar 

  15. 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

    Article  PubMed  CAS  Google Scholar 

  16. Lu W, Bennett BD, Rabinowitz JD (2008) Analytical strategies for LC–MS-based targeted metabolomics. J Chromatogr B 871:236–242

    Article  CAS  Google Scholar 

  17. 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

    Article  PubMed  CAS  Google Scholar 

  18. 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

    Article  CAS  Google Scholar 

  19. 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

    Article  PubMed  CAS  Google Scholar 

Download references

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

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints 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

Publish with us

Policies and ethics