Protocol for Using the HTX MALDI Matrix Sublimator for Mass Spec Imaging Sample Preparation v1

This protocol describes the standardised operation of the HTX Sublimator for automated MALDI matrix deposition by sublimation for mass spectrometry imaging (MSI) sample preparation. It outlines instrument setup, vacuum and temperature control, matrix loading, sublimation parameters, chamber flushing, and cleaning procedures to ensure reproducible and contamination-free matrix deposition. When performed as described, this protocol produces uniform and controlled matrix layers suitable for high-quality MSI experiments, with stable vacuum performance and minimal matrix carryover. This validated version includes recommended starting parameters for commonly used MALDI matrices and documents the limitation of automated recrystallisation, with guidance for performing recrystallisation manually when required.