Figure 1 from Determining the N-Glycan and Collagen/Extracellular Matrix Protein Compositions in a Novel Outcome Cohort of Prostate Cancer Tissue Microarrays Using MALDI-MSI

Workflow protocols for sample preparation. For N-glycan analysis, FFPE slides were deparaffinized and rehydrated before undergoing antigen retrieval in citraconic buffer. PNGaseF (1) was applied as a “dry” molecular coating before a 2-hour incubation at 37°C. CHCA matrix was applied, and imaging of N-glycans was performed. For analysis of core- vs outer-arm fucosylated N-glycans, enzyme EndoF3 (2) was dialyzed before application, and imaging of EndoF3-digested N-glycans was performed before subsequent PNGaseF digestion and imaging of remaining N-glycans. Following glycan imaging, CHCA matrix was removed from tissues, and samples were histologically analyzed following H&E staining. Tissues underwent a second antigen retrieval with tris buffer before COLaseIII (3) was applied as a “dry” molecular coating. Slides were incubated for 5 hours at 37°C. CHCA matrix was applied, and imaging of collagen/ECM peptides was performed.