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- Multiplex data utilizing a single-step staining and imaging workflow for the investigation of multiple sample types

# Multiplex data utilizing a single-step staining and imaging workflow for the investigation of multiple sample types

Resolving tissue complexity across whole slides has been limited by image resolution, the number of targets that can be assessed at once, and sample throughput; this AACR 2024 poster reports what one round of staining and one Orion scan produced across eight tissue examples, from breast and lung cancer to inflammatory bowel disease and type 1 diabetes.

Presented by RareCyte.

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- Every marker in the panel was stained in one round and imaged in one scan. Each section carried a 15- to 18-plex protein biomarker panel, and 20 immunofluorescence channels were collected and quantitated simultaneously — the protein targets, nuclear markers, and the tissue’s own autofluorescence isolated as a separate channel to add morphological detail — at a reported scan speed of around 70 minutes per square centimeter for 20 IF channels.

- H&E was then run on the very same section, with no serial sections required. Because the one-round immunofluorescence process preserved sample integrity, slides were de-coverslipped in aqueous solution, stripped of IF reagents and H&E stained, then imaged at 20X on the same Orion platform in brightfield mode, so morphology and quantitative spatial data could be aligned and overlaid across the same cells.

- Eight examples spanned healthy and diseased tissue across several organs. Figures 1–8 cover breast ductal adenocarcinoma, mouse ileum, non-small cell lung cancer, hepatocellular carcinoma, inflammatory bowel disease, and pancreas including type 1 diabetes; Figures 1 and 2 show 15-plex whole-slide panels beside the H&E of the same section.

[Download poster](https://cdn.sanity.io/files/s3p6uqps/production/7247dafad9b2e856904b401c6584cc22f7e7c727.pdf)

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[Download the poster](https://cdn.sanity.io/files/s3p6uqps/production/7247dafad9b2e856904b401c6584cc22f7e7c727.pdf) to view it.

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