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. 2020 Mar 3;4(5):708–723. doi: 10.1002/hep4.1494

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© 2020 The Authors. Hepatology Communications published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Spectral Library generated from optimized monoplex stains and an unstained liver biopsy section. A single slide from each antibody‐Opal fluorophore pair, a single slide with DAPI alone, and an unstained slide were analyzed using inForm software (included with the Vectra 3 platform) to create a “Spectral Library.” The Spectral Library is one of the main advantages of using the Vectra 3 platform, as it eliminates spectral overlap by unmixing the various fluorophore signals and allows subtraction of autofluorescence. Abbreviations: FITC, fluorescein isothiocyanate; TRITC, tetramethyl rhodamine isothiocyanate.