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Updated: May 2, 2026
High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence
Published on: October 28, 2025
Multiplex Immunofluorescence Staining of Paraffin-Embedded Tissue: Coverslip Mounting of Tissue Sections Improves
Gitte A Pedersen1, Søren H Elsborg1, Rikke Nørregaard1
1Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
Abstract:
Immunofluorescence (IF) microscopy is essential for studying protein expression and localization. Multiplex IF enables sequential staining of the same tissue section for comprehensive analysis but often involves harsh treatments or expensive reagents that can damage tissues and reduce signal quality. Here, we present a detailed protocol for multiplex IF using indirect antibody labeling, a mild stripping buffer, and cost-effective reagents. Importantly, mounting paraffin-embedded tissue sections on gelatin-coated coverslips enhances Z-axis optical resolution in widefield imaging.
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