Skip to main content
SpringerLink
Log in

The management of haemoglobin interference for the MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies

  • Communication
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

MALDI-MSI represents an ideal tool to explore the spatial distribution of proteins directly in situ, integrating molecular and cytomorphological information, enabling the discovery of potential diagnostic markers in thyroid cytopathology. However, red cells present in the fine needle aspiration biopsy (FNAB) specimens caused ion suppression of other proteins during the MALDI-MSI analysis due to large amount of haemoglobin. Aim of this study was to set up a sample preparation workflow able to manage this haemoglobin interference. Three protocols were compared using ex vivo cytological samples collected from fresh thyroid nodules of 9 patients who underwent thyroidectomy: (A) conventional air-dried smears, (B) cytological smears immediately fixed in ethanol, and (C) ThinPrep liquid-based preparation. Protocols C and A were also evaluated using real FNABs. Results show that protocol C markedly decreased the amount of haemoglobin, with respect to protocols A and B. Protein profiles obtained with protocols A and B were characterised by high inter-patient variability, probably related to the abundance of the haemoglobin, whereas similar spectra were observed for protocol C, where haemoglobin contents were lower. Our findings suggest protocol C as the sample preparation method for MALDI-MSI analysis.

Graphical abstract

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

Abbreviations

FNAB:

Fine needle aspiration biopsies

Hb:

Haemoglobin

H&E:

Haematoxylin and eosin

ITO:

Indium tin oxide

LBP:

Liquid-based preparation

MALDI:

Matrix-assisted laser desorption/ionisation

MSI:

Mass spectrometry imaging

ROI:

Region of interest

S/N:

Signal to noise

TIC:

Total ion current

References

  1. Melany M, Chen S. Thyroid cancer: ultrasound imaging and fine-needle aspiration biopsy. Endocrinol Metab Clin N Am. 2017;46:691–711. https://doi.org/10.1016/j.ecl.2017.04.011.

    Article  Google Scholar 

  2. Seeley EH, Caprioli RM. Molecular imaging of proteins in tissues by mass spectrometry. PNAS. 2008;105:18126–31. https://doi.org/10.1073/pnas.0801374105.

    Article  PubMed  Google Scholar 

  3. Mosele N, Smith A, Galli M, Pagni F, Magni F. MALDI-MSI analysis of cytological smears: the study of thyroid Cancer. In: Cole LM, editor. Imaging mass spectrometry. New York: Springer New York; 2017. p. 37–47. https://doi.org/10.1007/978-1-4939-7051-3_5.

    Chapter  Google Scholar 

  4. Schwamborn K, Krieg RC, Uhlig S, Ikenberg H, Wellmann H. MALDI imaging as a specific diagnostic tool for routine cervical cytology specimens. Int J Mol Med. 2011;27. https://doi.org/10.3892/ijmm.2010.587.

  5. Pagni F, De Sio G, Garancini M, Scardilli M, Chinello C, Smith AJ, et al. Proteomics in thyroid cytopathology: relevance of MALDI-imaging in distinguishing malignant from benign lesions. Proteomics. 2016;16:1775–84. https://doi.org/10.1002/pmic.201500448.

    Article  CAS  PubMed  Google Scholar 

  6. Mainini V, Pagni F, Garancini M, Giardini V, De Sio G, Cusi C, et al. An alternative approach in endocrine pathology research: MALDI-IMS in papillary thyroid carcinoma. Endocr Pathol. 2013;24:250–3. https://doi.org/10.1007/s12022-013-9273-8.

    Article  PubMed  Google Scholar 

  7. Pagni F, Mainini V, Garancini M, Bono F, Vanzati A, Giardini V, et al. Proteomics for the diagnosis of thyroid lesions: preliminary report. Cytopathology. 2015;26:318–24. https://doi.org/10.1111/cyt.12166.

    Article  CAS  PubMed  Google Scholar 

  8. Amann JM, Chaurand P, Gonzalez A, Mombley J, Massion P, Carbone D, et al. Selective profiling of proteins in lung cancer cells from fine-needle aspirates by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Canc Res. 2006;12:5142–50. https://doi.org/10.1158/1078-0432.CCR-06-0264.

    Article  CAS  Google Scholar 

  9. Howitt BE, Chang S, Eszlinger M, Paschke R, Drage MG, Krane JF, et al. Fine-needle aspiration diagnoses of noninvasive follicular variant of papillary thyroid carcinoma. Am J Clin Pathol. 2015;144:850–7. https://doi.org/10.1309/AJCPEIE12POICULI.

    Article  CAS  PubMed  Google Scholar 

  10. Linder J. Recent advances in thin-layer cytology. Diagn Cytopathol. 1998;18:24–32. https://doi.org/10.1002/(SICI)1097-0339(199801)18:1<24::AID-DC5>3.0.CO;2-U.

    Article  CAS  PubMed  Google Scholar 

  11. Ardito G, Rossi ED, Revelli L, Moschella F, Giustozzi E, Fadda G, et al. The role of fine-needle aspiration performed with liquid-based cytology in the surgical management of thyroid lesions. In Vivo. 2010;24:333–7.

    PubMed  Google Scholar 

  12. Smith A, Piga I, Galli M, Stella M, Denti V, Del Puppo M, et al. Matrix-assisted laser desorption/ionisation mass spectrometry imaging in the study of gastric cancer: a mini review. Int J Mol Sci. 2017;18(12):2588. https://doi.org/10.3390/ijms18122588.

    Article  CAS  PubMed Central  Google Scholar 

  13. Piga I, Heijs B, Nicolardi S, Giusti L, Marselli L, Marchetti P, et al. Ultra-high resolution MALDI-FTICR-MSI analysis of intact proteins in mouse and human pancreas tissue. Int J Mass Spectrom. 2019;437:10–6. https://doi.org/10.1016/j.ijms.2017.11.001.

    Article  CAS  Google Scholar 

  14. Gibb S, Strimmer K. MALDIquant: a versatile R package for the analysis of mass spectrometry data. Bioinformatics. 2012;28:2270–1. https://doi.org/10.1093/bioinformatics/bts447.

    Article  CAS  PubMed  Google Scholar 

  15. Zhang LM, Freitas MA, Wickham J, Parthun MR, Klisovic MI, Marcucci G, et al. Differential expression of histone post-translational modifications in acute myeloid and chronic lymphocytic leukemia determined by high-pressure liquid chromatography and mass spectrometry. J Am Soc Mass Spectrom. 2004;15(1):77–86. https://doi.org/10.1016/j.jasms.2003.10.001.

    Article  CAS  PubMed  Google Scholar 

  16. Michael CW, Hunter B. Interpretation of fine-needle aspirates processed by the ThinPrep technique: cytologic artifacts and diagnostic pitfalls. Diagn Cytopathol. 2000;23:6–13. https://doi.org/10.1002/1097-0339(200007)23:1<6::AID-DC2>3.0.CO;2-F.

    Article  CAS  PubMed  Google Scholar 

  17. Rossi ED, Zannoni GF, Moncelsi S, Stigliano E, Santeusanio G, Lombardi CP, et al. Application of liquid-based cytology to fine-needle aspiration biopsies of the thyroid gland. Front Endocrinol. 2012;3:57. https://doi.org/10.3389/fendo.2012.00057.

    Article  Google Scholar 

  18. Keyhani E, Sharghi SA, Amini R, Sharghi SA, Karimlou M, Moghaddam FA, et al. Liquid base cytology in evaluation of thyroid nodules. J Diabetes Metab Disord. 2014;13:82. https://doi.org/10.1186/s40200-014-0082-5.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Chong Y, Ji S-J, Kang CS, Lee EJ. Can liquid-based preparation substitute for conventional smear in thyroid fine-needle aspiration? A systematic review based on meta-analysis. Endocr Connect. 2017;6:817–29. https://doi.org/10.1530/EC-17-0165.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was funded by AIRC (Associazione Italiana per la Ricerca sul Cancro) MFAG GRANT 2016 - Id. 18445, and partially supported by the Italian Ministry of University and Research (MIUR) - Department of Excellence project PREMIA (PREcision MedIcine Approach: bringing biomarker research to clinic).

Author information

Author notes

  1. Isabella Piga and Giulia Capitoli contributed equally to this work

Authors and Affiliations

  1. School of Medicine and Surgery, Clinical Proteomics and Metabolomics Unit, University of Milano – Bicocca, 20854, Vedano al Lambro, Italy

    Isabella Piga, Vanna Denti, Silvia Tettamanti, Andrew Smith, Martina Stella, Clizia Chinello & Fulvio Magni

  2. School of Medicine and Surgery, Section of Pathology, University of Milano - Bicocca, Via Cadore 48, 20900, Monza, Italy

    Isabella Piga & Fabio Pagni

  3. School of Medicine and Surgery, Centre of Biostatistics for Clinical Epidemiology, University of Milano – Bicocca, 20854, Vedano al Lambro, Italy

    Giulia Capitoli & Stefania Galimberti

  4. Department of Radiology, San Gerardo Hospital, 20900, Monza, Italy

    Davide Leni

  5. Department of Surgery, San Gerardo Hospital, 20900, Monza, Italy

    Mattia Garancini

Authors
  1. Isabella Piga
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Giulia Capitoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vanna Denti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Silvia Tettamanti
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Andrew Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Martina Stella
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Clizia Chinello
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Davide Leni
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mattia Garancini
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Stefania Galimberti
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fulvio Magni
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Fabio Pagni
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fabio Pagni.

Ethics declarations

The study was approved by the local Ethical Board – San Gerardo Hospital – AIRC-MFAG 18/11/2016. Signed informed consent of all individual participant samples was obtained.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(PDF 711 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Piga, I., Capitoli, G., Denti, V. et al. The management of haemoglobin interference for the MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies. Anal Bioanal Chem 411, 5007–5012 (2019). https://doi.org/10.1007/s00216-019-01908-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-019-01908-w

Keywords

Search

Navigation