Sample preparation of formalin-fixed paraffin-embedded tissue sections for MALDI-mass spectrometry imaging

Juliane Hermann, Heidi Noels, Wendy Theelen, Michaela Lellig, Setareh Orth-Alampour, Peter Boor, Vera Jankowski, Joachim Jankowski*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MALDI MSI) has become a powerful tool with a high potential relevance for the analysis of biomolecules in tissue samples in the context of diseases like cancer and cardiovascular or cardiorenal diseases. In recent years, significant progress has been made in the technology of MALDI MSI. However, a more systematic optimization of sample preparation would likely achieve an increase in the molecular information derived from MALDI MSI. Therefore, we have employed a systematic approach to develop, establish and validate an optimized "standard operating protocol" (SOP) for sample preparation in MALDI MSI of formalin-fixed paraffin-embedded (FFPE) tissue sample analyses within this study. The optimized parameters regarding the impact on the resulting signal-to-noise (S/N) ratio were as follows: (i) trypsin concentration, solvents, deposition method, and incubation time; (ii) tissue washing procedures and drying processes; and (iii) spray flow rate, number of layers of trypsin deposition, and grid size. The protocol was evaluated on interday variability and its applicability for analyzing the mouse kidney, aorta, and heart FFPE tissue samples. In conclusion, an optimized SOP for MALDI MSI of FFPE tissue sections was developed to generate high sensitivity, to enhance spatial resolution and reproducibility, and to increase its applicability for various tissue types. This optimized SOP will further increase the molecular information content and intensify the use of MSI in future basic research and diagnostic applications.

Graphical

Original languageEnglish
Pages (from-to)1263-1275
Number of pages13
JournalAnalytical and Bioanalytical Chemistry
Volume412
Issue number6
DOIs
Publication statusPublished - Feb 2020

Keywords

  • MALDI imaging
  • Tissue preparation
  • Formalin-fixed paraffin-embedded tissue sections
  • Optimized sample standard operating protocols (SOP)
  • HIGH-RESOLUTION
  • PROTEOMIC ANALYSIS
  • PROTEINS
  • MSI
  • METASTASIS
  • PROTOCOLS
  • LIPIDS

Fingerprint

Dive into the research topics of 'Sample preparation of formalin-fixed paraffin-embedded tissue sections for MALDI-mass spectrometry imaging'. Together they form a unique fingerprint.

Cite this