Skip to main content
SpringerLink
Log in

Analysis of Lipids in Single Cells and Organelles Using Nanomanipulation-Coupled Mass Spectrometry

  • Protocol
  • First Online:
Single Cell Metabolism

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2064))

Abstract

The ability to discriminately analyze the chemical constituents of single cells and organelles is highly sought after and necessary to establish true biomarkers. Some major challenges of individual cell analysis include requirement and expenditure of a large sample of cells as well as extensive extraction and separation techniques. Here, we describe methods to perform individual cell and organelle extractions of both tissues and cells in vitro using nanomanipulation coupled to mass spectrometry. Lipid profiles display heterogeneity from extracted adipocytes and lipid droplets, demonstrating the necessity for single cell analysis. The application of these techniques can be applied to other cell and organelle types for selective and thorough monitoring of disease progression and biomarker discovery.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 199.99
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

Institutional subscriptions

References

  1. Teo CC, Chong WPK, Tan E, Basri NB, Low ZJ, Ho YS (2015) Advances in sample preparation and analytical techniques for lipidomics study of clinical samples. Trends Anal Chem 66:1–18

    Article  CAS  Google Scholar 

  2. Wenk MR (2005) The emerging field of lipidomics. Nat Rev Drug Discov 4:594–610

    Article  CAS  Google Scholar 

  3. Oikawa A, Saito K (2012) Metabolite analyses of single cells. Plant J 70:30–38

    Article  CAS  Google Scholar 

  4. Trouillon R, Passarelli MK, Wang J, Kurczy ME, Ewing AG (2013) Chemical analysis of single cells. Anal Chem 85:522–542

    Article  CAS  Google Scholar 

  5. Borland LM, Kottegoda S, Phillips KS, Allbritton NL (2008) Chemical analysis of single cells. Annu Rev Anal Chem 1:191–227

    Article  CAS  Google Scholar 

  6. Rubakhin SS, Lanni EJ, Sweedler JV (2013) Progress toward single cell metabolomics. Curr Opin Biotechnol 24:95–104

    Article  CAS  Google Scholar 

  7. Zenobi R (2013) Single-cell metabolomics: analytical and biological perspectives. Science 342:1201

    Article  CAS  Google Scholar 

  8. Ide Y, Waki M, Ishizaki I, Nagata Y, Yamazaki F, Hayasaka T, Masaki N, Ikegami K, Kondo T, Shibata K, Ogura H, Sanada N, Setou M (2014) Single cell lipidomics of SKBR-3 breast cancer cells by using time-of-flight secondary-ion mass spectrometry. Surf Interface Anal 46:181–184

    Article  CAS  Google Scholar 

  9. Speicher MR (2013) Single-cell analysis: toward the clinic. Genome Med 5:74/71–74/73. 73 pp

    Article  Google Scholar 

  10. Espina V, Wulfkuhle JD, Calvert VS, Van Meter A, Zhou W, Coukos G, Geho DH, Petricoin EF III, Liotta LA (2006) Laser capture microdissection. Nat Protoc 1:586–603

    Article  CAS  Google Scholar 

  11. Masujima T (2009) Live single-cell mass spectrometry. Anal Sci 25:953–960

    Article  CAS  Google Scholar 

  12. Ellis SR, Ferris CJ, Gilmore KJ, Mitchell TW, Blanksby SJ, in het Panhuis M (2012) Direct lipid profiling of single cells from inkjet printed microarrays. Anal Chem 84:9679–9683

    Article  CAS  Google Scholar 

  13. Lanni EJ, Rubakhin SS, Sweedler JV (2012) Mass spectrometry imaging and profiling of single cells. J Proteome 75:5036–5051

    Article  CAS  Google Scholar 

  14. Rubakhin SS, Garden RW, Fuller RR, Sweedler JV (2000) Measuring the peptides in individual organelles with mass spectrometry. Nat Biotechnol 18:172–175

    Article  CAS  Google Scholar 

  15. Chandra S, Lorey DR II (2001) SIMS ion microscopy in cancer research: single cell isotopic imaging for chemical composition, cytotoxicity and cell cycle recognition. Cell Mol Biol 47:503–518

    CAS  PubMed  Google Scholar 

  16. Whittal RM, Keller BO, Li L (1998) Nanoliter chemistry combined with mass spectrometry for peptide mapping of proteins from single mammalian cell lysates. Anal Chem 70:5344–5347

    Article  CAS  Google Scholar 

  17. Hebbar S, Schulz WD, Sauer U, Schwudke D (2014) Laser capture microdissection coupled with on-column extraction LC-MSn enables lipidomics of fluorescently labeled drosophila neurons. Anal Chem 86:5345–5352

    Article  CAS  Google Scholar 

  18. Wu H, Volponi JV, Oliver AE, Parikh AN, Simmons BA, Singh S (2011) In vivo lipidomics using single-cell raman spectroscopy. Proc Natl Acad Sci U S A 108:3809–3814, S3809/3801-S3809/3811

    Article  CAS  Google Scholar 

  19. Aerts JT, Louis KR, Crandall SR, Govindaiah G, Cox CL, Sweedler JV (2014) Patch clamp electrophysiology and capillary electrophoresis-mass spectrometry metabolomics for single cell characterization. Anal Chem 86:3203–3208

    Article  CAS  Google Scholar 

  20. Garden RW, Shippy SA, Li L, Moroz TP, Sweedler JV (1998) Proteolytic processing of the Aplysia egg-laying hormone prohormone. Proc Natl Acad Sci U S A 95:3972–3977

    Article  CAS  Google Scholar 

  21. Lorenzo TM, Mizuno H, Tsuyama N, Harada T, Masujima T (2012) In situ molecular analysis of plant tissues by live single-cell mass spectrometry. Anal Chem 84:5221–5228

    Article  Google Scholar 

  22. Horn PJ, Ledbetter NR, James CN, Hoffman WD, Case CR, Verbeck GF, Chapman KD (2011) Visualization of lipid droplet composition by direct organelle mass spectrometry. J Biol Chem 286:3298–3306

    Article  CAS  Google Scholar 

  23. Phelps M, Hamilton J, Verbeck GF (2014) Nanomanipulation-coupled nanospray mass spectrometry as an approach for single cell analysis. Rev Sci Instrum 85:124101–124105

    Article  Google Scholar 

  24. Phelps MS, Verbeck GF (2015) A lipidomics demonstration of the importance of single cell analysis. Anal Methods 7:3668–3670

    Article  CAS  Google Scholar 

  25. Phelps MS, Sturtevant D, Chapman KD, Verbeck G (2016) Nanomanipulation-Coupled Matrix-Assisted Laser Desorption/Ionization-Direct Organelle Mass Spectrometry: A Technique for the Detailed Analysis of Single Organelles. J Am Soc Mass Spectrom 27:187–193

    Article  CAS  Google Scholar 

  26. Huynh V, Joshi U, Leveille JM, Golden TD, Verbeck GF (2014) Nanomanipulation-coupled to nanospray mass spectrometry applied to document and ink analysis. Forensic Sci Int 242:150–156

    Article  CAS  Google Scholar 

  27. Clemons K, Wiley R, Waverka K, Fox J, Dziekonski E, Verbeck GF (2013) Direct analyte-probed nanoextraction coupled to nanospray ionization-mass spectrometry of drug residues from latent fingerprints. J Forensic Sci 58:875–880

    Article  CAS  Google Scholar 

  28. Ledbetter NL, Walton BL, Davila P, Hoffmann WD, Ernest RN, Verbeck GFIV (2010) Nanomanipulation-coupled nanospray mass spectrometry applied to the extraction and analysis of trace analytes found on fibers. J Forensic Sci 55:1218–1221

    Article  CAS  Google Scholar 

  29. Clemons K, Dake J, Sisco E, GFT V (2013) Trace analysis of energetic materials via direct analyte-probed nanoextraction coupled to direct analysis in real time mass spectrometry. Forensic Sci Int 231:98–101

    Article  CAS  Google Scholar 

  30. Lysy PA, Smets F, Sibille C, Najimi M, Sokal EM (2007) Human skin fibroblasts: from mesodermal to hepatocyte-like differentiation. Hepatology 46:1574–1585

    Article  CAS  Google Scholar 

  31. Madsen L, Petersen RK, Kristiansen K (2005) Regulation of adipocyte differentiation and function by polyunsaturated fatty acids. Biochim Biophys Acta Mol basis Dis 1740:266–286

    Article  CAS  Google Scholar 

  32. Ntambi JM, Kim Y-C (2000) Adipocyte differentiation and gene expression. J Nutr 130:3122S–3126S

    Article  CAS  Google Scholar 

  33. Bruins AP (1991) Mass spectrometry with ion sources operating at atmospheric pressure. Mass Spectrom Rev 10:53–77

    Article  CAS  Google Scholar 

  34. Cazes J (2004) Analytical instrumentation handbook. CRC Press, Boca Raton

    Book  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. University of North Texas, Denton, TX, USA

    Mandy S. Phelps & Guido F. Verbeck

Authors
  1. Mandy S. Phelps
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guido F. Verbeck
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guido F. Verbeck .

Editor information

Editors and Affiliations

  1. Waters Corporation, Milford, MA, USA

    Bindesh Shrestha

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Phelps, M.S., Verbeck, G.F. (2020). Analysis of Lipids in Single Cells and Organelles Using Nanomanipulation-Coupled Mass Spectrometry. In: Shrestha, B. (eds) Single Cell Metabolism. Methods in Molecular Biology, vol 2064. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9831-9_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9831-9_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9829-6

  • Online ISBN: 978-1-4939-9831-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation