Multimodal Imaging of the Cellular and Extracellular Microenvironment on the Same Formalin-Fixed Paraffin-Embedded Tissue Section
Same-tissue section multimodal imaging is a powerful strategy that spatially profiles tissue histology, cell populations and molecular composition while maximizing tissue economy, preserving spatial molecular relationships, and increasing co-registration capacity. However, performing multiple modalities on the same tissue section can destroy or chemically alter the tissue, compromising downstream…
Multitask imaging of tissue sections has the potential to provide a detailed understanding of both tissue structure and molecular composition. However, conducting multiple imaging techniques on the same tissue can damage the sample or alter its chemical properties, impacting the quality of the resulting data. This study systematically investigates the integration of three different staining methods - picrosirius red, hematoxylin and eosin, and multiplexed immunofluorescence - together with two IMS workflows that analyze N-glycan and extracellular matrix peptide composition.
By evaluating the impact of these integrations on tissue morphology, stain efficiency, IMS feature intensity and localization, the researchers propose a sequence that maximizes data quality. The optimized workflow begins with autofluorescence microscopy, followed by multiplexed immunofluorescence, picrosirius red staining, N-glycan IMS, hematoxylin and eosin staining, and finally extracellular matrix peptide IMS.
This new methodology allows for the comprehensive imaging of tissue morphology, collagen fibers, individual cell populations, N-glycan composition, and extracellular matrix peptides at single-cell resolution from a single, undamaged tissue section.
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