Skip to main content
SpringerLink
Log in

Identification of oligosaccharides from histopathological sections by MALDI imaging mass spectrometry

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Direct tissue analysis using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) provides the means for in situ molecular analysis of a wide variety of biomolecules. This technology—known as imaging mass spectrometry (IMS)—allows the measurement of biomolecules in their native biological environments without the need for target-specific reagents such as antibodies. In this study, we applied the IMS technique to formalin-fixed paraffin-embedded samples to identify a substance(s) responsible for the intestinal obstruction caused by an unidentified foreign body. In advance of IMS analysis, some pretreatments were applied. After the deparaffinization of sections, samples were subjected to enzyme digestion. The sections co-crystallized with matrix were desorbed and ionized by a laser pulse with scanning. A combination of α-amylase digestion and the 2,5-dihydroxybenzoic acid matrix gave the best mass spectrum. With the IMS Convolution software which we developed, we could automatically extract meaningful signals from the IMS datasets. The representative peak values were m/z 1,013, 1,175, 1,337, 1,499, 1,661, 1,823, and 1,985. Thus, it was revealed that the material was polymer with a 162-Da unit size, calculated from the even intervals. In comparison with the mass spectra of the histopathological specimen and authentic materials, the main component coincided with amylopectin rather than amylose. Tandem MS analysis proved that the main components were oligosaccharides. Finally, we confirmed the identification of amylopectin by staining with periodic acid-Schiff and iodine. These results for the first time show the advantages of MALDI-IMS in combination with enzyme digestion for the direct analysis of oligosaccharides as a major component of histopathological samples.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

DHB:

2,5-Dihydroxybenzoic acid

FFPE:

Formalin-fixed paraffin-embedded

HE:

Hematoxylin and eosin

IMS:

Imaging mass spectrometry

ITO:

Indium tin oxide

MALDI:

Matrix-assisted laser desorption/ionization

PAS:

Periodic acid-Schiff stain

ROI:

Region of interest

CT:

Computed tomography

References

  1. Yao I, Sugiura Y, Matsumoto M, Setou M (2008) In situ proteomics with imaging mass spectrometry and principal component analysis in the Scrapper-knockout mouse brain. Proteomics 8(18):3692–3701

    Article  CAS  Google Scholar 

  2. Sugiura Y, Setou M (2010) Imaging mass spectrometry for visualization of drug and endogenous metabolite distribution: toward in situ pharmacometabolomes. J Neuroimmune Pharmacol 5(1):31–43. doi:10.1007/s11481-009-9162-6

    Article  Google Scholar 

  3. Bernier UR, Kline DL, Barnard DR, Schreck CE, Yost RA (2000) Analysis of human skin emanations by gas chromatography/mass spectrometry. 2. Identification of volatile compounds that are candidate attractants for the yellow fever mosquito (Aedes aegypti). Anal Chem 72(4):747–756

    Article  CAS  Google Scholar 

  4. Garrett TJ, Yost RA (2006) Analysis of intact tissue by intermediate-pressure MALDI on a linear ion trap mass spectrometer. Anal Chem 78(7):2465–2469

    Article  CAS  Google Scholar 

  5. Drexler DM, Garrett TJ, Cantone JL, Diters RW, Mitroka JG, Prieto Conaway MC, Adams SP, Yost RA, Sanders M (2007) Utility of imaging mass spectrometry (IMS) by matrix-assisted laser desorption ionization (MALDI) on an ion trap mass spectrometer in the analysis of drugs and metabolites in biological tissues. J Pharmacol Toxicol Methods 55(3):279–288

    Article  CAS  Google Scholar 

  6. Shimma S, Sugiura Y, Hayasaka T, Hoshikawa Y, Noda T, Setou M (2007) MALDI-based imaging mass spectrometry revealed abnormal distribution of phospholipids in colon cancer liver metastasis. J Chromatogr 855(1):98–103

    Article  CAS  Google Scholar 

  7. 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(4):493–496

    Article  CAS  Google Scholar 

  8. Setou M, Heeren RM, Stoeckli M, Simma S, Matsumoto M (2007) Mass microscopy. Seikagaku 79(9):874–879

    CAS  Google Scholar 

  9. Rubakhin SS, Churchill JD, Greenough WT, Sweedler JV (2006) Profiling signaling peptides in single mammalian cells using mass spectrometry. Anal Chem 78(20):7267–7272

    Article  CAS  Google Scholar 

  10. Luxembourg SL, Mize TH, McDonnell LA, Heeren RM (2004) High-spatial resolution mass spectrometric imaging of peptide and protein distributions on a surface. Anal Chem 76(18):5339–5344

    Article  CAS  Google Scholar 

  11. Cooks RG, Ouyang Z, Takats Z, Wiseman JM (2006) Detection technologies. Ambient mass spectrometry. Science (New York, NY) 311(5767):1566–1570

    Article  CAS  Google Scholar 

  12. Sugiura Y, Shimma S, Setou M (2006) Two-step matrix application technique to improve ionization efficiency for matrix-assisted laser desorption/ionization in imaging mass spectrometry. Anal Chem 78(24):8227–8235

    Article  CAS  Google Scholar 

  13. Shimma S, Sugiura Y, Hayasaka T, Zaima N, Matsumoto M, Setou M (2008) Mass imaging and identification of biomolecules with MALDI-QIT-TOF-based system. Anal Chem 80(3):878–885

    Article  CAS  Google Scholar 

  14. Landgraf RR, Garrett TJ, Calcutt NA, Stacpoole PW, Yost RA (2007) MALDI-linear ion trap microprobe MS/MS studies of the effects of dichloroacetate on lipid content of nerve tissue. Anal Chem 79(21):8170–8175

    Article  CAS  Google Scholar 

  15. Shimma S, Furuta M, Ichimura K, Yoshida Y, Setou M (2006) A novel approach to in situ proteome analysis using a chemical inkjet printing technology and MALDI-QIT-TOF tandem mass spectrometer. J Mass Spectrom Soc Japan 54:133–140

    Article  CAS  Google Scholar 

  16. Groseclose MR, Andersson M, Hardesty WM, Caprioli RM (2007) Identification of proteins directly from tissue: in situ tryptic digestions coupled with imaging mass spectrometry. J Mass Spectrom 42(2):254–262

    Article  CAS  Google Scholar 

  17. Walch A, Rauser S, Deininger SO, Hofler H (2008) MALDI imaging mass spectrometry for direct tissue analysis: a new frontier for molecular histology. Histochem Cell Biol 130(3):421–434

    Article  CAS  Google Scholar 

  18. 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):4751–4760

    Article  CAS  Google Scholar 

  19. Morita Y, Ikegami K, Goto-Inoue N, Hayasaka T, Zaima N, Tanaka H, Uehara T, Setoguchi T, Sakaguchi T, Igarashi H, Sugimura H, Setou M, Konno H (2009) Imaging mass spectrometry of gastric carcinoma in formalin-fixed paraffin-embedded tissue microarray. Cancer Sci 101(1):267–273

    Article  Google Scholar 

  20. Ushijima M, Miyata S, Eguchi S, Kawakita M, Yoshimoto M, Iwase T, Akiyama F, Sakamoto G, Nagasaki K, Miki Y, Noda T, Hoshikawa Y, Matsuura M (2007) Common peak approach using mass spectrometry data sets for predicting the effects of anticancer drugs on breast cancer. Cancer Inform 3:285–293

    Google Scholar 

  21. Gahrton G (1964) Microspectrophotometric quantitation of the periodic acid-Schiff (PAS) reaction in human neutrophil leukocytes based on a model system of glycogen microdroplets. Exp Cell Res 34:488–506

    Article  CAS  Google Scholar 

  22. Holness CL, Simmons DL (1993) Molecular cloning of CD68, a human macrophage marker related to lysosomal glycoproteins. Blood 81(6):1607–1613

    CAS  Google Scholar 

  23. Hayasaka T, Goto-Inoue N, Ushijima M, Yao I, Yuba-Kubo A, Wakui M, Kajihara S, Matsuura M, Setou M (2011) Development of imaging mass spectrometry (IMS) dataset extractor software, IMS convolution. Anal Bioanal Chem 401:183–193

    Article  CAS  Google Scholar 

  24. Domon B, Costello CE (1988) Structure elucidation of glycosphingolipids and gangliosides using high-performance tandem mass spectrometry. Biochemistry 27(5):1534–1543

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Hosaka K, Saitou A, and Tarumi T for their assistance. We also thank Prof. Sugimura and lab members. This study is supported by Research Grants for PRESTO and SENTAN from Japan Science and Technology Agency to I. Yao.

Author information

Authors and Affiliations

  1. Department of Surgery, Kansai Medical University, 2-3-1 Shin-machi, Hirakata, Osaka, 573-1191, Japan

    Masanori Yamada, Hideho Takada & A-Hon Kwon

  2. Department of Medical Chemistry, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi, Osaka, 570-8506, Japan

    Ikuko Yao & Seiji Ito

  3. Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 7 Goban-cho, Chiyoda-ku, Tokyo, 102-0076, Japan

    Ikuko Yao

  4. Department of Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan

    Takahiro Hayasaka & Mitsutoshi Setou

  5. Bioinformatics Group, Genome Center, Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto-ku, Tokyo, 135-8550, Japan

    Masaru Ushijima & Masaaki Matsuura

  6. Division of Cancer Genomics, Cancer Institute, Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto-ku, Tokyo, 135-8550, Japan

    Masaaki Matsuura

  7. Department of Clinical Sciences and Laboratory Medicine, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi, Osaka, 570-8506, Japan

    Nobuaki Shikata

Authors
  1. Masanori Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ikuko Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takahiro Hayasaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Masaru Ushijima
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masaaki Matsuura
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hideho Takada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nobuaki Shikata
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Mitsutoshi Setou
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A-Hon Kwon
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Seiji Ito
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ikuko Yao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yamada, M., Yao, I., Hayasaka, T. et al. Identification of oligosaccharides from histopathological sections by MALDI imaging mass spectrometry. Anal Bioanal Chem 402, 1921–1930 (2012). https://doi.org/10.1007/s00216-011-5622-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-011-5622-y

Keywords

Search

Navigation