Hematoxylin and Eosin staining of PhenoCycler® Fusion flow cell slides

ABSTRACT

Multiplexed Imaging technologies are powerful techniques that enable ultrahigh-plex spatial phenotyping of whole tissue sections at single cell spatial resolution. Co-Detection by Indexing (CODEX) multiplexing can detect up to 100 proteins using cyclic detection of DNA conjugated antibodies applied to tissue sections. However, it is necessary to correlate multiplexed fluorescent (mIF) spatial images with Hematoxylin and Eosin (H&E) stained sections post analysis. To effectively correlate mIF spatial images with H&E morphology, an (H&E) staining protocol was developed that is directly applied to the CODEX Fusion flow-cell slide after analysis allowing for direct H&E correlation and annotation with mIF images.

KEYWORDS:

• H&E
• PhenoCyler® Fusion
• flow cell

Acknowledgements

The authors thank Cynthia Loomis and Shanmugapriya Selvaraj of the NYU Langone Health Experimental Pathology Laboratory for their expertise and guidance. We thank Boris Reizis for his guidance and financial support (AE). We thank Sylvia Adams and Natalie Klar of the NYU Perlmutter Cancer Center at NYU Langone Health for providing breast cancer specimens.

Ethics disclosure

This work was approved by the NYU Langone Health Institutional Review Board.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported in part by the NYULH Pathology Department Translational Research Program (TRP) and by the NIH Grant AI128949. The NYULH Center for Biospecimen Research and Development, Histology and Immunohistochemistry Laboratory (RRID:SCR_018304) is supported in part by the Laura and Isaac Perlmutter Cancer Center Support Grant; NIH/NCI P30CA016087. AE received funding from the German Research Foundation DFG (EI1185/1-1).