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Gelatin-coated indium tin oxide slides improve human cartilage-bone tissue adherence and N-glycan signal intensity for mass spectrometry imaging

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Abstract

Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) has been successfully used to elucidate the relative abundance and spatial mapping of analytes in situ. Currently, sample preparation workflows for soft formalin-fixed paraffin-embedded (FFPE) tissues, such as brain, liver, kidney, and heart, have been successfully developed. However, hard tissues, such as cartilage-bone, tooth, and whole mouse body, have resulted in the loss of morphology or tissue during the heat-induced epitope retrieval (HIER) step on commercially available conductive indium tin oxide (ITO) slides. Therefore, we have successfully developed a novel and cost-effective sample preparation workflow in which commercial conductive ITO slides are pre-coated with gelatin and chromium potassium sulfate dodecahydrate to improve the adherence of FFPE human osteoarthritic cartilage-bone tissue sections. Gelatin-coated ITO slides also resulted in overall higher N-glycan signal intensity for not only FFPE osteoarthritic cartilage-bone tissue but also for FFPE hard-boiled egg white used as a quality control to assess the quality of sample preparation and MALDI-MSI acquisition. In summary, we present a novel straightforward workflow to improve slide adherence and morphological preservation of FFPE cartilage-bone tissue sections during HIER while improving the signal intensity of N-glycans spatially mapped from the same tissue sections by MALDI-MSI.

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Abbreviations

CACH:

α-Cyano-4-hydroxycinnamic acid

EDTA:

Ethylenediaminetetraacetic acid

FF:

Fresh-frozen

FFPE:

Formalin-fixed paraffin-embedded

HIER:

Heat-induced epitope retrieval

ITO:

Indium tin oxide

LC-ESI MS/MS:

Liquid chromatography electrospray ionisation mass spectrometry

MALDI-MSI:

Matrix-assisted laser desorption/ionisation mass spectrometry imaging

RMS:

Root mean square

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Acknowledgments

The authors acknowledge Bioplatforms Australia, the University of South Australia, and the State and Federal Governments, which co-fund the NCRIS-enabled Mass Spectrometry and Proteomics facility at the University of South Australia. The authors also wish to thank Dr. Dzenita Muratovic for providing a human tibial plateau specimen, and the University of South Australia for Postgraduate Award and Research Training Program domestic fee offset scholarship.

Funding

This research was funded by Dr. Malcolm Cochran.

Author information

Authors and Affiliations

  1. Clinical and Health Sciences, Health and Biomedical Innovation, University of South Australia, Adelaide, SA, 5001, Australia

    Yea-Rin Lee & Paul H. Anderson

  2. Future Industries Institute, Mawson Lakes Campus, University of South Australia, Mawson Lakes, SA, 5095, Australia

    Yea-Rin Lee, Matthew T. Briggs, Mark R. Condina & Peter Hoffmann

  3. Discipline of Orthopaedics and Trauma, Adelaide Medical School, The University of Adelaide, Adelaide, SA, 5000, Australia

    Yea-Rin Lee & Julia S. Kuliwaba

Authors
  1. Yea-Rin Lee
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  2. Matthew T. Briggs
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  3. Julia S. Kuliwaba
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  4. Paul H. Anderson
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  5. Mark R. Condina
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  6. Peter Hoffmann
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Contributions

Conceptualisation, Y-R. Lee, MT. Briggs, JS. Kuliwaba, PH. Anderson, MR. Condina, and P. Hoffmann; methodology, Y-R. Lee and MT. Briggs; data analysis, Y-R. Lee and MT. Briggs; writing—original draft preparation, Y-R. Lee; writing—review and editing, MT. Briggs, JS. Kuliwaba, PH. Anderson, MR. Condina, and P. Hoffmann; supervision, MT. Briggs, JS. Kuliwaba, PH. Anderson, and P. Hoffman; funding acquisition, PH. Anderson. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Peter Hoffmann.

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The authors declare that they have no conflict of interest.

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Published in the topical collection Mass Spectrometry Imaging 2.0 with guest editors Shane R. Ellis and Tiffany Porta Siegel.

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Lee, YR., Briggs, M.T., Kuliwaba, J.S. et al. Gelatin-coated indium tin oxide slides improve human cartilage-bone tissue adherence and N-glycan signal intensity for mass spectrometry imaging. Anal Bioanal Chem 413, 2675–2682 (2021). https://doi.org/10.1007/s00216-020-02986-x

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