ArticleSTAR protocols2026
Protocol for extended-plex immunofluorescence staining of FFPE tissues using a sequential bleach-and-stain approach.
Article in STAR protocols, 2026. 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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7 authors.
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Abstract
Multiplex immunofluorescence enables spatial analysis of complex biological systems, but increasing plex levels raise cost and workflow complexity. Here, we present a protocol for extending tyramide signal amplification (TSA)-based multiplex immunofluorescence from 6 to 16 markers using a sequential bleach-and-stain approach. It covers tissue preparation, panel establishment, sequential staining, fluorophore bleaching, image acquisition and computational image processing including stitching, registration, and overlay. The workflow is applicable to formalin-fixed paraffin-embedded (FFPE) human cancer and lymphoid tissues and is adaptable through the selection of appropriate antibodies.
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