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Water-assisted laser desorption/ionization mass spectrometry for minimally invasive in vivo and real-time surface analysis using SpiderMass

Nature Protocols volume 14, pages 3162–3182 (2019)Cite this article

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Abstract

Rapid, sensitive, precise and accurate analysis of samples in their native in vivo environment is critical to better decipher physiological and physiopathological mechanisms. SpiderMass is an ambient mass spectrometry (MS) system designed for mobile in vivo and real-time surface analyses of biological tissues. The system uses a fibered laser, which is tuned to excite the most intense vibrational band of water, resulting in a process termed water-assisted laser desorption/ionization (WALDI). The water molecules act as an endogenous matrix in a matrix-assisted laser desorption ionization (MALDI)-like scenario, leading to the desorption/ionization of biomolecules (lipids, metabolites and proteins). The ejected material is transferred to the mass spectrometer through an atmospheric interface and a transfer line that is several meters long. Here, we formulate a three-stage procedure that includes (i) a laser system setup coupled to a Waters Q-TOF or Thermo Fisher Q Exactive mass analyzer, (ii) analysis of specimens and (iii) data processing. We also describe the optimal setup for the analysis of cell cultures, fresh-frozen tissue sections and in vivo experiments on skin. With proper optimization, the system can be used for a variety of different targets and applications. The entire procedure takes 1–2 d for complex samples.

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Fig. 1: A schematic representation of the SpiderMass setup for in vivo experiments.
Fig. 2: A flowchart of a standard SpiderMass experiment.
Fig. 3: Close-up of the microsampling probe.
Fig. 4: Interface.
Fig. 5: Typical MS spectra of bovine liver.
Fig. 6: Reproducibility experiment.
Fig. 7: Two-class classification of 33 dog sarcoma samples.

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Data availability

All published data presented here are publicly available and can be found in the PRIDE Archive (https://www.ebi.ac.uk/pride/archive/). The dog sarcoma data are available from the ProteomeXchange Consortium (PXD010990), Real-time molecular diagnosis of tumors using water-assisted laser desorption/ionization mass spectrometry technology.

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Acknowledgements

We thank all members of SATT-Nord involved in SpiderMass and, in particular, E. Rollet and F.-X. Denimal for their support of the project. This work was funded by the Ministère de l’Enseignement Supérieur, de la Recherche et de l’Innovation, Université de Lille and Inserm. The project was also funded by ANR-14-CE17-0021 REALITY’MS (I.F.), Inserm Programme PhysiCancer SPIDERMASS (M.S.), Région Hauts de France-EU FEDER O’DREAMS (D.T., I.F. and M.S.) and SIRIC ONCOLille grant INCa-DGOS-Inserm 6041aa (D.T., I.F. and M.S.). SpiderMass was awarded Best Breakthrough Innovation Prize by the MATWIN 2015 international board.

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Author notes

  1. These authors contributed equally: Nina Ogrinc, Philippe Saudemont.

Authors and Affiliations

  1. Université de Lille, Inserm U1192, Laboratoire Protéomique, Réponse Inflammatoire et Spectrométrie de Masse (PRISM), Villeneuve d’Ascq, France

    Nina Ogrinc, Philippe Saudemont, Yves-Marie Robin, Jean-Pascal Gimeno, Quentin Pascal, Dominique Tierny, Michel Salzet & Isabelle Fournier

  2. SATT-Nord, Immeuble Central Gare, Lille, France

    Philippe Saudemont

  3. Department of Surgery and Cancer, St Mary’s Hospital, Imperial College London, London, UK

    Julia Balog & Zoltan Takats

  4. Unité de Pathologie Morphologique et Moléculaire, Centre Oscar Lambret, Lille, France

    Yves-Marie Robin

  5. OCR (Oncovet Clinical Research), Eurasanté, Loos, France

    Quentin Pascal & Dominique Tierny

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Contributions

N.O., P.S. and J.B. wrote the original draft of the manuscript. P.S., N.O. and J.-P.G. carried out the experiments. P.S. and I.F. developed the technique. Q.P. collected the samples and performed the histology. Y.-M.R. did histology and validated diagnostics. P.S., J.B. and I.F. analyzed the data. I.F., M.S. and D.T. designed the study. I.F., M.S. and Z.T. corrected the manuscript. I.F. and M.S. supervised the project and provided the funding.

Corresponding authors

Correspondence to Michel Salzet or Isabelle Fournier.

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Competing interests

J.B. is an employee of Waters Research Center. M.S. and I.F. are inventors on a patent (priority no. WO2015IB57301 20150922) related to part of the described protocol. Q.P. is an employee of OCR. D.T. is founder and CEO of OCR. Z.T. is a consultant for Waters Corporation. The system is under protection by patent CA2961491 A1 (29). The remaining authors declare no competing interests.

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Key references using this protocol

Fatou, B. et al. Sci. Rep. 6, 25919 (2016): https://www.nature.com/articles/srep25919

Fatou, B. et al. J. Biotechnol. 281, 61–66 (2018): https://www.sciencedirect.com/science/article/pii/S0168165618305169

Saudemont, P. et al. Cancer Cell 34, 840–851.E4 (2018): https://www.cell.com/cancer-cell/fulltext/S1535-6108(18)30423-9

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Ogrinc, N., Saudemont, P., Balog, J. et al. Water-assisted laser desorption/ionization mass spectrometry for minimally invasive in vivo and real-time surface analysis using SpiderMass. Nat Protoc 14, 3162–3182 (2019). https://doi.org/10.1038/s41596-019-0217-8

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