Analytical and Bioanalytical Chemistry

, Volume 407, Issue 8, pp 2159–2166 | Cite as

Increasing specificity in imaging mass spectrometry: high spatial fidelity transfer of proteins from tissue sections to functionalized surfaces

Research Paper
Part of the following topical collections:
  1. Mass Spectrometry Imaging


Imaging mass spectrometry (IMS) is a technique in full expansion used in many clinical and biological applications. A common limitation of the technology, particularly true for protein analysis, is that only the most abundant and/or more easily ionizable molecules are typically detected. One approach to overcome this limitation is to transfer proteins contained within tissue sections onto functionalized surfaces with high spatial fidelity for IMS analysis. In this case, only proteins with an affinity for the surface will be retained whereas others will be removed. The chemical nature of the surface is therefore critical. The research work presented herein proposes a high spatial fidelity transfer method for proteins from thin tissue sections onto a nitrocellulose surface. The method employs a home-built apparatus that allows the transfer process to be performed without any direct physical contact between the section and the transfer surface while maintaining physical pressure between the surfaces to help protein migration. The performance of this system was demonstrated using mouse liver and kidney sections. Serials sections were also collected either to be stained with hematoxylin and eosin (H&E) to assess the spatial fidelity of the transfer process or to be directly analyzed as a control sample to differentiate the signals detected after transfer. IMS results showed a high spatial fidelity transfer of a subset of proteins. Some of the detected proteins were poorly observed or not observed with conventional direct tissue analysis, demonstrating an increase in detection sensitivity and specificity with the newly developed method.

Graphical Abstract

Imaging MS of proteins transferred from tissue sections to a capture membrane


Imaging mass spectrometry MALDI Proteins Nitrocellulose Transfer 

Supplementary material

216_2014_8300_MOESM1_ESM.pdf (8.7 mb)
ESM 1(PDF 8951 kb)


  1. 1.
    Cornett DS, Reyzer ML, Chaurand P, Caprioli RM (2007) MALDI imaging mass spectrometry: molecular snapshots of biochemical systems. Nat Meth 4(10):828–833CrossRefGoogle Scholar
  2. 2.
    Norris JL, Caprioli RM (2013) Analysis of tissue specimens by matrix-assisted laser desorption/ionization imaging mass spectrometry in biological and clinical research. Chem Rev 113(4):2309–2342CrossRefGoogle Scholar
  3. 3.
    Stoeckli M, Chaurand P, Hallahan DE, Caprioli RM (2001) Imaging mass spectrometry: a new technology for the analysis of protein expression in mammalian tissues. Nat Med 7:493–496CrossRefGoogle Scholar
  4. 4.
    Chughta K, Heeren RMA (2010) Mass spectrometric imaging for biomedical tissue analysis. Chem Rev 110:3237–3277CrossRefGoogle Scholar
  5. 5.
    Thomas A, Chaurand P (2014) Advances in tissue section preparation for MALDI imaging MS. Bioanalysis 6(7):967–982CrossRefGoogle Scholar
  6. 6.
    Chaurand P, Fouchecourt S, Dague BB, Xu BJ, Reyzer ML, Orgebin-Crist M-C, Caprioli RM (2003) Profiling and imaging proteins in the mouse epididymis by imaging mass spectrometry. Proteomics 3:2221–2239CrossRefGoogle Scholar
  7. 7.
    Chaurand P, Rahman MA, Hunt T, Mobley JA, Gu G, Lathan JC, Caprioli RM, Kasper S (2008) Monitoring mouse prostate development by profiling and imaging mass spectrometry. Mol Cell Proteomics 7:411–423CrossRefGoogle Scholar
  8. 8.
    Chaurand P, Sanders ME, Jensen RA, Caprioli RM (2004) Proteomics in diagnostic pathology: profiling and imaging proteins directly in tissue sections. Am J Pathol 165:1057–1068CrossRefGoogle Scholar
  9. 9.
    Norris JL, Caprioli RM (2013) Imaging mass spectrometry: a new tool for pathology in a molecular age. Proteomics Clin Appl 7(11–12):733–738CrossRefGoogle Scholar
  10. 10.
    Schwamborn K, Caprioli RM (2010) MALDI imaging mass spectrometry—painting molecular pictures. Mol Oncol 4(6):529–538CrossRefGoogle Scholar
  11. 11.
    Reyzer ML, Caldwell RL, Dugger TC, Forbes JT, Ritter CA, Guix M, Arteaga CL, Caprioli RM (2004) Early changes in protein expression detected by mass spectrometry predict tumor response to molecular therapeutics. Cancer Res 64(24)Google Scholar
  12. 12.
    Reyzer ML, Caprioli RM (2007) MALDI-MS-based imaging of small molecules and proteins in tissues. Curr Opin Chem Biol 11(1):29–35CrossRefGoogle Scholar
  13. 13.
    Chaurand P, Norris JL, Cornett DS, Mobley JA, Caprioli RM (2006) New developments in profiling and imaging of proteins from tissue sections by MALDI mass spectrometry. J Proteome Res 5:2889–2900CrossRefGoogle Scholar
  14. 14.
    Thomas A, Laveaux Charbonneau J, Fournaise E, Chaurand P (2012) Sublimation of new matrix candidates for high spatial resolution imaging mass spectrometry of lipids: enhanced information in both positive and negative polarities after 1,5-diaminonaphthalene deposition. Anal Chem 84:2048–2054CrossRefGoogle Scholar
  15. 15.
    Dufresne M, Thomas A, Breault-Turcot J, Masson J-F, Chaurand P (2013) Silver assisted laser desorption ionization for high spatial resolution imaging mass spectrometry of olefins from thin tissue sections. Anal Chem 85:3318–3324CrossRefGoogle Scholar
  16. 16.
    Chaurand P, Caprioli RM (2002) Direct profiling and imaging of peptides and proteins from mammalian cells and tissue sections by mass spectrometry. Electrophoresis 23(18):3125–3135CrossRefGoogle Scholar
  17. 17.
    Liu R, Li Q, Smith LM (2014) Detection of large ions in time-of-flight mass spectrometry: effects of ion mass and acceleration voltage on microchannel plate detector response. J Am Soc Mass Spectrom 25(8):1374–1383CrossRefGoogle Scholar
  18. 18.
    Lemaire R, Stauber J, Wisztorski M, Van Camp C, Desmons A, Deschamps M, Proess G, Rudlof I, Woods AS, Day R, Salzet M, Fournier I (2007) Tag-mass: specific molecular imaging of transcriptome and proteome by mass spectrometry based on photocleavable tag. J Proteome Res 6(6):2057–2067CrossRefGoogle Scholar
  19. 19.
    Thiery G, Anselmi E, Audebourg A, Darii E, Abarbri M, Terris B, Tabet J-C, Gut IG (2008) Improvements of TArgeted multiplex mass spectrometry IMaging. Proteomics 8(18):3725–3734CrossRefGoogle Scholar
  20. 20.
    Thiery G, Shchepinov MS, Southern EM, Audebourg A, Audard V, Terris B, Gut IG (2007) Multiplex target protein imaging in tissue sections by mass spectrometry—TAMSIM. Rapid Commun Mass Spectrom 21(6):823–829CrossRefGoogle Scholar
  21. 21.
    Yang J, Chaurand P, Norris JL, Porter NA, Caprioli RM (2012) Activity-based probes linked with laser-cleavable mass tags for signal amplification in imaging mass spectrometry: analysis of serine hydrolase enzymes in mammalian tissue. Anal Chem 84(8):3689–3695CrossRefGoogle Scholar
  22. 22.
    Caprioli RM, Farmer TB, Gile J (1997) Molecular imaging of biological samples: localization of peptides and proteins using MALDI-TOF MS. Anal Chem 69(23):4751–4760CrossRefGoogle Scholar
  23. 23.
    Chaurand P, Dague BB, Pearsall RS, Threadgill DW, Caprioli RM (2001) Profiling proteins from azoxymethane-induced colon tumors at the molecular level by MALDI mass spectrometry. Proteomics 1:1320–1326CrossRefGoogle Scholar
  24. 24.
    Chaurand P, Stoeckli M, Caprioli RM (1999) Direct profiling of proteins in biological tissue sections by maldi mass spectrometry. Anal Chem 71:5263–5270CrossRefGoogle Scholar
  25. 25.
    Bouamrani A, Ternier J, Ratel D, Benabid A-L, Issartel J-P, Brambilla E, Berger F (2006) Direct-tissue SELDI-TOF mass spectrometry analysis: a new application for clinical proteomics. Clin Chem 52:2103–2105CrossRefGoogle Scholar
  26. 26.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets—procedure and some applications. Proc Natl Acad Sci U S A 76(9):4350–4354CrossRefGoogle Scholar
  27. 27.
    Luque-Garcia JL, Zhou G, Sun T-T, Neubert TA (2006) Use of nitrocellulose membranes for protein characterization by matrix-assisted laser desorption/ionization mass spectrometry. Anal Chem 78(14):5102–5108CrossRefGoogle Scholar
  28. 28.
    Lobert S, Correia JJ (1994) Method for rapid electrophoretic transfer of isoelectric-focusing gels to polyvinylidene difluoride. Electrophoresis 15(7):930–931CrossRefGoogle Scholar
  29. 29.
    Chaurand P, Cornett DS, Caprioli RM (2006) Molecular imaging of thin mammalian tissue sections by mass spectrometry. Curr Opin Biotechnol 17(4):431–436CrossRefGoogle Scholar
  30. 30.
    Tang N, Tornatore P, Weinberger SR (2004) Current developments in SELDI affinity technology. Mass Spectrom Rev 23:34–44CrossRefGoogle Scholar
  31. 31.
    Müller M, Gras R, Appel RD, Bienvenut WV, Hochstrasser DF (2002) Visualization and analysis of molecular scanner peptide mass spectra. J Am Soc Mass Spectrom 13(3):221–231CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Department of ChemistryUniversité de MontréalMontréalCanada

Personalised recommendations