Visualization of Small Intact Proteins in Breast Cancer FFPE Tissue

Methods Mol Biol. 2023:2688:161-172. doi: 10.1007/978-1-0716-3319-9_14.

Abstract

Molecular visualization of metabolites, lipids, and proteins by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) is becoming an in-demand analytical approach to aid the histopathological analysis of breast cancer. Particularly, proteins seem to play a role in cancer progression, and specific proteins are currently used in the clinic for staging. Formalin-fixed paraffin-embedded (FFPE) tissues are ideal for correlating the molecular markers with clinical outcomes due to their long-term storage. So far, to obtain proteomic information by MSI from this kind of tissue, antigen retrieval and tryptic digestion steps are required. In this chapter, we present a protocol to spatially detect small proteins in tumor and necrotic regions of patient-derived breast cancer xenograft FFPE tissues without employing any on-tissue digestion. This protocol can be used for other kinds of FFPE tissue following specific optimization of the sample preparation phases.

Keywords: Breast cancer; FFPE tissue; MALDI MSI; Protein.

MeSH terms

  • Breast Neoplasms*
  • Female
  • Formaldehyde / chemistry
  • Humans
  • Paraffin Embedding
  • Proteins
  • Proteomics / methods
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
  • Tissue Fixation / methods

Substances

  • Proteins
  • Formaldehyde