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.

Piga, I., Capitoli, G., Denti, V., Tettamanti, S., Smith, A., Stella, M., et al. (2019). The management of haemoglobin interference for the MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 411(20), 5007-5012 [10.1007/s00216-019-01908-w].

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

Piga, I;Capitoli, G;Denti, V;Smith, A;Stella, M;Chinello, C;Garancini, M;Galimberti, S;Magni, F;Pagni, F
2019

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.
No
Articolo in rivista - Articolo scientifico
Scientifica
Fine needle aspiration; MALDI mass spectrometry imaging; proteomics; Haemoglobin interference; Thyroid; ThinPrep
English
5007
5012
6
Piga, I., Capitoli, G., Denti, V., Tettamanti, S., Smith, A., Stella, M., et al. (2019). The management of haemoglobin interference for the MALDI-MSI proteomics analysis of thyroid fine needle aspiration biopsies. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 411(20), 5007-5012 [10.1007/s00216-019-01908-w].
Piga, I; Capitoli, G; Denti, V; Tettamanti, S; Smith, A; Stella, M; Chinello, C; Leni, D; Garancini, M; Galimberti, S; Magni, F; Pagni, F
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/10281/231167
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