ArticlebioRxiv : the preprint server for biology2025
Protocol for spatial characterization of ECM collagen-GAG in CRC tumor microenvironment.
Article in bioRxiv : the preprint server for biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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8 authors.
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Abstract
The extracellular matrix (ECM) plays a critical role in colorectal cancer (CRC) progression and therapeutic resistance. Accurate characterization of ECM composition and architecture is essential for understanding how CRC evades therapy, yet most protocols either assess ECM components in isolation or remain technically challenging. Here we present a robust yet simple protocol for spatial characterization of collagen and glycosaminoglycan (GAG) organization within the CRC tumor microenvironment (TME). Our method combines Alcian Blue and Picrosirius Red staining procedures with standardized tissue processing and imaging protocols. The protocol enables simultaneous visualization, assessment, and quantification of collagen and GAG distribution patterns in formalin-fixed, paraffin-embedded tissue sections. Key methodological advances include optimized dual-staining approach with distinct blue-red coloration for straightforward spectral separation on digital imaging systems, standardized reagent preparations, and validated imaging parameters. The complementary wavelengths facilitate both visual interpretation and potential digital separation, offering advantages over multi-component stains with overlapping spectral ranges. Validation across multiple CRC patient specimens demonstrates excellent reproducibility with consistent staining intensity using standard histology equipment. Because the resulting spatial maps can be compared directly with engineered or
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