MALDI Direct Analysis and Imaging of Frozen Versus FFPE Tissues: What Strategy for Which Sample?

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Mass Spectrometry Imaging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 656))

Abstract

Significant advances have been made in the past decade in the field of mass spectrometry imaging with MALDI ion sources (MALDI-MSI). While MALDI-MSI has high potential in the field of biology and in the clinic, a challenge for MALDI-MSI has been to adapt itself to a greater range of sample types. In particular, much of the biological archived materials for pathology studies are tissue biopsies fixed with paraformaldehyde and embedded in paraffin (FFPE tissues) because of the high stability of such samples. Thus, there has been a need to develop strategies for analyzing FFPE samples as this would allow retrospective studies of past clinical cases on large cohorts of existing samples. Obviously, PAF fixation, by inducing protein cross-linking, causes problems for molecular analysis by MS. We developed on tissue digestion strategies for overcoming these difficulties and allowing molecular data to be retrieved from FFPE samples no matter how long they have been stored. These digestion strategies preserve localization from digested proteins making MALDI-MSI of proteins possible by monitoring the resulting peptides. We present methods and protocols for FFPE samples. These strategies have proven to be valuable for all tested FFPE samples and have opened archived tissues from hospital banks to MALDI-MSI.

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References

  1. Reyzer, M. L., Hsieh, Y., Ng, K., Korfmacher, W. A., Caprioli, R. M. (2003) Direct analysis of drug candidates in tissue by matrix-assisted laser desorption/ionization mass spectrometry. J Mass Spectrom, 38, 1081–1092.

    Article  CAS  PubMed  Google Scholar 

  2. Rubakhin, S. S., Jurchen, J. C., Monroe, E. B., Sweedler, J. V. (2005) Imaging mass spectrometry: fundamentals and applications to drug discovery. Drug Discov Today, 10, 823–837.

    Article  CAS  PubMed  Google Scholar 

  3. Deininger, S. O., Ebert, M. P., Futterer, A., Gerhard, M., Rocken, C. (2008) MALDI imaging combined with hierarchical clustering as a new tool for the interpretation of complex human cancers. J Proteome Res, 7, 5230–5236

    Article  CAS  PubMed  Google Scholar 

  4. Fournier, I., Wisztorski, M., Salzet, M. (2008) Tissue imaging using MALDI-MS: a new frontier of histopathology proteomics. Expert Rev Proteomics, 5, 413–424.

    Article  CAS  PubMed  Google Scholar 

  5. Franck, J., Arafah, K., Elayed, M., Bonnel, D., Vergara, D., Jacquet, A., Vinatier, D., Wisztorski, M., Day, R., Fournier, I., Salzet, M. (2009) MALDI IMAGING: state of the art technology in clinical proteomics. Mol Cell Proteomics, 8, 2023-2033.

    Article  CAS  PubMed  Google Scholar 

  6. Walch, A., Rauser, S., Deininger, S. O., Hofler, H. (2008) MALDI imaging mass spectrometry for direct tissue analysis: a new frontier for molecular histology. Histochem Cell Biol, 130, 421–434.

    Article  CAS  PubMed  Google Scholar 

  7. Groseclose, M. R., Andersson, M., Hardesty, W. M., Caprioli, R. M. (2007) Identification of proteins directly from tissue: in situ tryptic digestions coupled with imaging mass spectrometry. J Mass Spectrom, 42, 254–262.

    Article  CAS  PubMed  Google Scholar 

  8. Lemaire, R., Desmons, A., Tabet, J. C., Day, R., Salzet, M., Fournier, I. (2007) Direct analysis and MALDI imaging of formalin-fixed, paraffin-embedded tissue sections. J Proteome Res, 6, 1295–1305.

    Article  CAS  PubMed  Google Scholar 

  9. Franck, J., Wisztorski, m., El ayed, M., Bonnel, D., Barnes, A., Fournier, I., Salzet, M. (2008) Automatic spotting solution for MALDI Imaging: process optimization and new developments. 56th Annual Conference on Mass Spectrometry and Allied Topics, Denver, CO, June 1–5, 2008.

    Google Scholar 

  10. Stauber, J., Lemaire, R., Franck, J., Bonnel, D., Croix, D., Day, R., Wisztorski, M., Fournier, I., Salzet, M. (2008) MALDI imaging of formalin-fixed paraffin-embedded tissues: application to model animals of Parkinson disease for biomarker hunting. J Proteome Res, 7, 969–978.

    Article  CAS  PubMed  Google Scholar 

  11. Groseclose, M. R., Massion, P. P., Chaurand, P., Caprioli, R. M. (2008) High-throughput proteomic analysis of formalin-fixed paraffin-embedded tissue microarrays using MALDI imaging mass spectrometry. Proteomics, 8, 3715–3724.

    Article  CAS  PubMed  Google Scholar 

  12. Ronci, M., Bonanno, E., Colantoni, A., Pieroni, L., Di Ilio, C., Spagnoli, L. G., Federici, G., Urbani, A. (2008) Protein unlocking procedures of formalin-fixed paraffin-embedded tissues: application to MALDI-TOF imaging MS investigations. Proteomics, 8, 3702–3714.

    Article  CAS  PubMed  Google Scholar 

  13. Plenat, F., Antunes, L., Haller, T., Piet-Ounnoughene, M., Klein-Monhoven, N., Champigneulle, J., Chenal, P., Bland, V., Garcia-Pimenta, F., Labouyrie, E. (2001) Formaldehyde fixation in the third millennium. Ann Pathol, 21, 29–47.

    CAS  PubMed  Google Scholar 

  14. Plenat, F., Montagne, K., Weinbreck, N., Corby, S., Champigneulle, J., Antunes, L., Bonnet, C., Maire, C., Monhoven, N. (2006) Molecular consequences of fixation and tissue processing: the examples of nucleic acids and proteins. Ann Pathol, 26, 8–21.

    Article  PubMed  Google Scholar 

  15. Kieman, J. A. (2000) Formaldehyde, formalin, paraformaldehyde and glutaraldehyde: what they are and what they do. Micros Today , 8, 8–14.

    Google Scholar 

  16. Redeker, V., Toullec, J. Y., Vinh, J., Rossier, J., Soyez, D. (1998) Combination of peptide profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and immunodetection on single glands or cells. Anal Chem, 70, 1805–1811.

    Article  CAS  PubMed  Google Scholar 

  17. Shi, S. R., Cote, R. J., Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: past, present, future. J Histochem Cytochem, 45, 327–343.

    Article  CAS  PubMed  Google Scholar 

  18. Shi, S. R., Cote, R. J., Taylor, C. R. (2001) Antigen retrieval techniques: current perspectives. J Histochem Cytochem, 49, 931–937.

    Article  CAS  PubMed  Google Scholar 

  19. Shi, S. R., Cote, R. J., Taylor, C. R. (2001) Antigen retrieval immunohistochemistry and molecular morphology in the year 2001. Appl Immunohistochem Mol Morphol, 9, 107–116.

    Article  CAS  PubMed  Google Scholar 

  20. Shi, S. R., Liu, C., Young, L., Taylor, C. (2007) Development of an optimal antigen retrieval protocol for immunohistochemistry of retinoblastoma protein (pRB) in formalin fixed, paraffin sections based on comparison of different methods. Biotech Histochem, 82, 301–309.

    Article  CAS  PubMed  Google Scholar 

  21. Shi, S. R., Shi, Y., Taylor, C. R. (2007) Updates on antigen retrieval techniques for immunohistochemistry. Chin J Pathol, 36, 7–10.

    CAS  Google Scholar 

  22. Taylor, C. R., Shi, S. R. (2000) Antigen retrieval: call for a return to first principles. Appl Immunohistochem Mol Morphol, 8, 173–174.

    Article  CAS  PubMed  Google Scholar 

  23. Xu, H., Yang, L., Wang, W., Shi, S. R., Liu, C., Liu, Y., Fang, X., Taylor, C. R., Lee, C. S., Balgley, B. M. (2008) Antigen retrieval for proteomic characterization of formalin-fixed and paraffin-embedded tissues. J Proteome Res, 7, 1098–1108.

    Article  CAS  PubMed  Google Scholar 

  24. Chaurand, P., Latham, J. C., Lane, K. B., Mobley, J. A., Polosukhin, V. V., Wirth, P. S., Nanney, L. B., Caprioli, R. M. (2008) Imaging mass spectrometry of intact proteins from alcohol-preserved tissue specimens: bypassing formalin fixation. J Proteome Res, 7, 3543–3555.

    Article  CAS  PubMed  Google Scholar 

  25. Lemaire, R., Tabet, J. C., Ducoroy, P., Hendra, J. B., Salzet, M., Fournier, I. (2006) Solid ionic matrixes for direct tissue analysis and MALDI imaging. Anal Chem, 78, 809–819.

    Article  CAS  PubMed  Google Scholar 

  26. Lemaire, R., Wisztorski, M., Desmons, A., Tabet, J. C., Day, R., Salzet, M., Fournier, I. (2006) MALDI-MS direct tissue analysis of proteins: improving signal sensitivity using organic treatments. Anal Chem, 78, 7145–7153.

    Article  CAS  PubMed  Google Scholar 

  27. Seeley, E. H., Oppenheimer, S. R., Mi, D., Chaurand, P., Caprioli, R. M. (2008) Enhancement of protein sensitivity for MALDI imaging mass spectrometry after chemical treatment of tissue sections. J Am Soc Mass Spectrom, 19, 1069–1077.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by grants from the Centre National de la Recherche Scientifique (DPI), Ministère de L’Enseignement Supérieur et de la Recherche, the Agence Nationale de la Recherche PCV (To IF).

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Wisztorski, M., Franck, J., Salzet, M., Fournier, I. (2010). MALDI Direct Analysis and Imaging of Frozen Versus FFPE Tissues: What Strategy for Which Sample?. In: Rubakhin, S., Sweedler, J. (eds) Mass Spectrometry Imaging. Methods in Molecular Biology, vol 656. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-746-4_18

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  • DOI: https://doi.org/10.1007/978-1-60761-746-4_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-745-7

  • Online ISBN: 978-1-60761-746-4

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