Cytological Cytospin Preparation for the Spatial Proteomics Analysis of Thyroid Nodules Using MALDI-MSI

Abstract

The application of innovative spatial omics approaches in the context of cytological specimens may open new frontiers for their diagnostic assessment. In particular, spatial proteomics using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) represents one of the most promising avenues, owing to its capability to map the distribution of hundreds of proteins within a complex cytological background in a multiplexed and relatively high-throughput manner. This approach may be particularly beneficial in the heterogeneous context of thyroid tumors where certain cells may not present clear-cut malignant morphology upon fine-needle aspiration biopsy, highlighting the necessity for additional molecular tools which are able to improve their diagnostic performance. This chapter aims to provide a detailed overview of a cytospin-based preparation workflow that has been optimized to facilitate the reliable spatial proteomics analysis of cytological thyroid specimens using MALDI-MSI, indicating the key aspects which should be considered when handling such samples.