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Confirmation of sub-cellular resolution using oversampling imaging mass spectrometry

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Abstract

The use of oversampling in MALDI (matrix-assisted laser desorption/ionization) imaging mass spectrometry (IMS) to improve lateral resolution is a common practice. However, its application is still controversial and recent studies reported a spot size–dependent change in the relative intensity of the spectra. Previously, using oversampling, we described the lipidome of the human colon epithelium, a 20–30 μm wide cell monolayer; even assessing the changes occurring within this monolayer associated with complex biological processes. Interestingly, the K-means analysis of those experiments unveiled the presence of a third epithelial cluster that anatomically matched the nuclei position. Taking into account the nucleus size (9–12 μm of diameter) and its distinctive lipidome, we decided to test whether this cluster was really of nuclear origin. Hence, the spectra obtained directly from tissue sections were compared with those recorded from the nuclei isolated from colon biopsies. The highest correlation coefficient was obtained when comparing the spectrum of the isolated nuclei with that of the tissue nuclear cluster, demonstrating the successful identification of the nuclear lipidome in the MALDI-IMS experiments run using oversampling and a lateral resolution of 10 μm/pixel. Importantly, it was established that phosphatidylinositol 38:4 nuclear levels remained stable along the colon crypt. That is, it mimicked neither the regular decrease observed in the epithelium nor the regular increase observed in the stroma, eliminating the chance of inter-pixel contamination. Altogether, besides confirming the usefulness of the oversampling technique, these results strongly reinforce the pivotal role IMS may have in promising fields such as single-cell analysis.

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References

  1. Baker TC, Han J, Borchers CH. Recent advancements in matrix-assisted laser desorption/ionization mass spectrometry imaging. Curr Opin Biotechnol. 2017;43:62–9. https://doi.org/10.1016/j.copbio.2016.09.003.

    Article  CAS  PubMed  Google Scholar 

  2. Oetjen J, Lachmund D, Palmer A, Alexandrov T, Becker M, Boskamp T, et al. An approach to optimize sample preparation for MALDI imaging MS of FFPE sections using fractional factorial design of experiments. Anal Bioanal Chem. 2016;408:6729–40. https://doi.org/10.1007/s00216-016-9793-4.

    Article  CAS  PubMed  Google Scholar 

  3. Swales JG, Dexter A, Hamm G, Nilsson A, Strittmatter N, Michopoulos F, et al. Quantitation of endogenous metabolites in mouse tumors using mass-spectrometry imaging. Anal Chem. 2018;90:6051–8. https://doi.org/10.1021/acs.analchem.7b05239.

    Article  CAS  PubMed  Google Scholar 

  4. Schwamborn K, Kriegsmann M, Weichert W. MALDI imaging mass spectrometry — from bench to bedside. Biochim Biophys Acta Proteins Proteom. 2017;1865:776–83. https://doi.org/10.1016/j.bbapap.2016.10.014.

    Article  CAS  PubMed  Google Scholar 

  5. Alexandrov T. MALDI imaging mass spectrometry: statistical data analysis and current computational challenges. BMC Bioinformatics. 2012;13:S11. https://doi.org/10.1186/1471-2105-13-S16-S11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Jurchen JC, Rubakhin SS, Sweedler JV. MALDI-MS imaging of features smaller than the size of the laser beam. J Am Soc Mass Spectrom. 2005;16:1654–9. https://doi.org/10.1016/j.jasms.2005.06.006.

    Article  CAS  PubMed  Google Scholar 

  7. Garate J, Fernández R, Lage S, Bestard-Escalas J, Lopez DH, Reigada R, et al. Imaging mass spectrometry increased resolution using 2-mercaptobenzothiazole and 2,5-diaminonaphtalene matrices: application to lipid distribution in human colon. Anal Bioanal Chem. 2015;407:4697–708. https://doi.org/10.1007/s00216-015-8673-7.

    Article  CAS  PubMed  Google Scholar 

  8. Lopez DH, Bestard-Escalas J, Garate J, Maimó-Barceló A, Fernández R, Reigada R, et al. Tissue-selective alteration of ethanolamine plasmalogen metabolism in dedifferentiated colon mucosa. Biochim Biophys Acta Mol Cell Biol Lipids. 2018;1863:928–38. https://doi.org/10.1016/j.bbalip.2018.04.017.

    Article  CAS  PubMed  Google Scholar 

  9. Bestard-Escalas J, Garate J, Maimó-Barceló A, Fernández R, Lopez DH, Lage S, et al. Lipid fingerprint image accurately conveys human colon cell pathophysiologic state: a solid candidate as biomarker. Biochim Biophys Acta Mol Cell Biol Lipids. 2016;1861:1942–50. https://doi.org/10.1016/j.bbalip.2016.09.013.

    Article  CAS  Google Scholar 

  10. Sparvero LJ, Amoscato AA, Dixon CE, Long JB, Kochanek PM, Pitt BR, et al. Mapping of phospholipids by MALDI imaging (MALDI-MSI): realities and expectations. Chem Phys Lipids. 2012;165:545–62. https://doi.org/10.1016/j.chemphyslip.2012.06.001.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Dufresne M, Guneysu D, Patterson NH, Marcinkiewicz MM, Regina A, Demeule M, et al. Multimodal detection of GM2 and GM3 lipid species in the brain of mucopolysaccharidosis type II mouse by serial imaging mass spectrometry and immunohistochemistry. Anal Bioanal Chem. 2017;409:1425–33. https://doi.org/10.1007/s00216-016-0076-x.

    Article  CAS  PubMed  Google Scholar 

  12. Duncan KD, Fang R, Yuan J, Chu RK, Dey SK, Burnum-Johnson KE, et al. Quantitative mass spectrometry imaging of prostaglandins as silver ion adducts with nanospray desorption electrospray ionization. Anal Chem. 2018;90:7246–52. https://doi.org/10.1021/acs.analchem.8b00350.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Cahill JF, Kertesz V, Van Berkel GJ. Characterization and application of a hybrid optical microscopy/laser ablation liquid vortex capture/electrospray ionization system for mass spectrometry imaging with sub-micrometer spatial resolution. Anal Chem. 2015;87:11113–21. https://doi.org/10.1021/acs.analchem.5b03293.

    Article  CAS  PubMed  Google Scholar 

  14. Korte AR, Yandeau-Nelson MD, Nikolau BJ, Lee YJ i. Subcellular-level resolution MALDI-MS imaging of maize leaf metabolites by MALDI-linear ion trap-orbitrap mass spectrometer. Anal Bioanal Chem. 2015;407:2301–9. https://doi.org/10.1007/s00216-015-8460-5.

    Article  CAS  PubMed  Google Scholar 

  15. Kertesz V, Van Berkel GJ. Improved imaging resolution in desorption electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom. 2008;22:2639–44. https://doi.org/10.1002/rcm.3662.

    Article  CAS  PubMed  Google Scholar 

  16. Bokhart MT, Manni J, Garrard KP, Ekelöf M, Nazari M, Muddiman DC. IR-MALDESI mass spectrometry imaging at 50 micron spatial resolution. J Am Soc Mass Spectrom. 2017. https://doi.org/10.1007/s13361-017-1740-x.

  17. Wiegelmann M, Dreisewerd K, Soltwisch J. Influence of the laser spot size, focal beam profile, and tissue type on the lipid signals obtained by MALDI-MS imaging in oversampling mode. J Am Soc Mass Spectrom. 2016;27:1952–64. https://doi.org/10.1007/s13361-016-1477-y.

    Article  CAS  PubMed  Google Scholar 

  18. Boggio KJ, Obasuyi E, Sugino K, Nelson SB, Agar NY, Agar JN. Recent advances in single-cell MALDI mass spectrometry imaging and potential clinical impact. Expert Rev Proteomics. 2011;8:591–604. https://doi.org/10.1586/epr.11.53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Fernández R, Carriel V, Lage S, Garate J, Díez-García J, Ochoa B, et al. Deciphering the lipid architecture of the rat sciatic nerve using imaging mass spectrometry. ACS Chem Neurosci. 2016;7:624–32. https://doi.org/10.1021/acschemneuro.6b00010.

    Article  CAS  PubMed  Google Scholar 

  20. Fernández R, González P, Lage S, Garate J, Maqueda A, Marcaida I, et al. Influence of the cation adducts in the analysis of matrix-assisted laser desorption ionization imaging mass spectrometry data from injury models of rat spinal cord. Anal Chem. 2017;89:8565–73. https://doi.org/10.1021/acs.analchem.7b02650.

    Article  CAS  PubMed  Google Scholar 

  21. Lagarrigue M, Becker M, Lavigne R, Deininger S-O, Walch A, Aubry F, et al. Revisiting rat spermatogenesis with MALDI imaging at 20-μm resolution. Mol Cell Proteomics. 2011;10:M110.005991. https://doi.org/10.1074/mcp.M110.005991.

    Article  CAS  PubMed  Google Scholar 

  22. Fernández R, Lage S, Abad-García B, Barceló-Coblijn G, Terés S, López DH, et al. Analysis of the lipidome of xenografts using MALDI-IMS and UHPLC-ESI-QTOF. J Am Soc Mass Spectrom. 2014;25:1237–46. https://doi.org/10.1007/s13361-014-0882-3.

    Article  CAS  PubMed  Google Scholar 

  23. Irvine RF. Nuclear lipid signalling. Nat Rev Mol Cell Biol. 2003;4:349–61. https://doi.org/10.1038/nrm1100.

    Article  CAS  PubMed  Google Scholar 

  24. Maté SM, Brenner RR, Ves-Losada A. Endonuclear lipids in liver cells This paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell. Can J Physiol Pharmacol. 2006;84:459–68. https://doi.org/10.1139/y05-097.

    Article  PubMed  Google Scholar 

  25. Ohsaki Y, Kawai T, Yoshikawa Y, Cheng J, Jokitalo E, Fujimoto T. PML isoform II plays a critical role in nuclear lipid droplet formation. J Cell Biol. 2016;212:29–38. https://doi.org/10.1083/jcb.201507122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Zhang L, Vertes A. Single-cell mass spectrometry approaches to explore cellular heterogeneity. Angew Chem Int Ed. 2018;57:4466–77. https://doi.org/10.1002/anie.201709719.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We are greatly thankful to the nurses and the medical doctors of the Gastroenterology Unit of Hospital Universitari de Son Espases (HUSE, Palma, Spain) and Hospital Comarcal d'Inca (Inca, Spain). Technical and human support provided by the Plataforma de Microscopía of IdISBa and by the Servicio de Lipidómica of the SGIKER (UPV/EHU, MICINN, GV/EJ, ESF) is gratefully acknowledged.

Funding

This study was funded by The Institute of Health Carlos III (CP12/03338), and the MINECO (Ministry of Economy and Competitiveness, RTC-2015-3693-1), and the Basque Government (IT1162-19) partially supported this study. The following institutions and projects supported authors’ contracts: AM-B, Institute of Health Carlos III (ISCIII, PI16/02200); JG, University of the Basque Country (UPV/EHU); JB-E, Govern Balear-Direcció General d'Innovació i Recerca (predoctoral fellowship, FPI/1787/2015); DHL and RF, Ministry of Economy and Competitiveness (RTC-2015-3693-1); and GB-C, ISCIII (Miguel Servet II program, CPII17/0005). Further, contracts and projects were co-funded either by the European Regional Development Fund (ERDF) or the European Social Fund (ESF).

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Author notes

  1. Roberto Fernández

    Present address: Department of Research and Development, IMG Pharma Biotech S.L., 48160, Derio, Spain

Authors and Affiliations

  1. Lipids in Human Pathology, Research Unit-Hospital Universitari Son Espases, Institut d’Investigació Sanitària Illes Balears (IdISBa) – Balearic Islands Health Research Institute, Ctra. Valldemossa 79, Module I, Floor -1, 07120, Palma, Balearic Islands, Spain

    Albert Maimó-Barceló, Joan Bestard-Escalas, Luzie Berthold, Daniel H. Lopez & Gwendolyn Barceló-Coblijn

  2. Department of Physical Chemistry, Fac. of Science and Technology, University of the Basque Country (UPV/EHU), Barrio Sarriena s/n, 48940, Leioa, Spain

    Jone Garate, Roberto Fernández & José Andrés Fernández

  3. Bioanalytical Department, Technical University of Berlin, Straße des 17. Juni 135, 10623, Berlin, Germany

    Luzie Berthold

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  1. Albert Maimó-Barceló
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Correspondence to José Andrés Fernández or Gwendolyn Barceló-Coblijn.

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Maimó-Barceló, A., Garate, J., Bestard-Escalas, J. et al. Confirmation of sub-cellular resolution using oversampling imaging mass spectrometry. Anal Bioanal Chem 411, 7935–7941 (2019). https://doi.org/10.1007/s00216-019-02212-3

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