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Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness
CONTEXT AND OBJECTIVE: The conversion of immunohistochemical (IHC) results from 3-dimensional tissue to a 2-dimensional visual image without considering tissue thickness poses a considerable risk of misleading IHC intensities. The present study aimed to clarify whether tissue thickness interferes wi...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Lippincott Williams & Wilkins
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7993914/ https://www.ncbi.nlm.nih.gov/pubmed/32404698 http://dx.doi.org/10.1097/PAI.0000000000000859 |
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author | Masuda, Shinobu Suzuki, Ryohei Kitano, Yuriko Nishimaki, Haruna Kobayashi, Hiroko Nakanishi, Yoko Yokoi, Hideo |
author_facet | Masuda, Shinobu Suzuki, Ryohei Kitano, Yuriko Nishimaki, Haruna Kobayashi, Hiroko Nakanishi, Yoko Yokoi, Hideo |
author_sort | Masuda, Shinobu |
collection | PubMed |
description | CONTEXT AND OBJECTIVE: The conversion of immunohistochemical (IHC) results from 3-dimensional tissue to a 2-dimensional visual image without considering tissue thickness poses a considerable risk of misleading IHC intensities. The present study aimed to clarify whether tissue thickness interferes with the estimation of IHC staining intensity and to introduce a control system to manage it. DESIGN: We prepared cell lines that are used as controls for human epidermal growth factor receptor 2 (HER2) IHC (MDA-MB-231, MDA-MB-175VII, MDA-MV-453, and SK-BR-3), a polyclonal antibody for HER2, an interferometry to measure the tissue thickness of formalin-fixed paraffin-embedded sections, a microscope with a Halogen or an LED light source, a complementary metal-oxide semiconductor camera in which the output signal can be corrected to γ=1, and a program to estimate color elements (hue, saturation, and luminance). It was examined whether tissue thickness interferes with the experimental scoring systems and practical classification of the routine HER2 scoring system. RESULTS: A noncellular control was shown to be better than a cellular control for managing tissue thickness. The IHC intensity for HER2 was correlated with tissue thickness (R(2)=0.8094), even under the less-standardized condition, but this correlation was better under the improved standardized condition using corrected γ=1 (R(2)=0.9282). Discrepancies in practical HER2 scores were increased in sections with thicknesses <2 and >5 μm. A control system to manage tissue thickness was introduced. CONCLUSIONS: Tissue thickness interferes with the estimation of the IHC intensity of HER2 in both experimental and practical scoring systems. A control system for managing tissue thickness is essential to increase the benefits of IHC as a standardized assay for clinical applications. |
format | Online Article Text |
id | pubmed-7993914 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Lippincott Williams & Wilkins |
record_format | MEDLINE/PubMed |
spelling | pubmed-79939142021-04-02 Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness Masuda, Shinobu Suzuki, Ryohei Kitano, Yuriko Nishimaki, Haruna Kobayashi, Hiroko Nakanishi, Yoko Yokoi, Hideo Appl Immunohistochem Mol Morphol Research Articles CONTEXT AND OBJECTIVE: The conversion of immunohistochemical (IHC) results from 3-dimensional tissue to a 2-dimensional visual image without considering tissue thickness poses a considerable risk of misleading IHC intensities. The present study aimed to clarify whether tissue thickness interferes with the estimation of IHC staining intensity and to introduce a control system to manage it. DESIGN: We prepared cell lines that are used as controls for human epidermal growth factor receptor 2 (HER2) IHC (MDA-MB-231, MDA-MB-175VII, MDA-MV-453, and SK-BR-3), a polyclonal antibody for HER2, an interferometry to measure the tissue thickness of formalin-fixed paraffin-embedded sections, a microscope with a Halogen or an LED light source, a complementary metal-oxide semiconductor camera in which the output signal can be corrected to γ=1, and a program to estimate color elements (hue, saturation, and luminance). It was examined whether tissue thickness interferes with the experimental scoring systems and practical classification of the routine HER2 scoring system. RESULTS: A noncellular control was shown to be better than a cellular control for managing tissue thickness. The IHC intensity for HER2 was correlated with tissue thickness (R(2)=0.8094), even under the less-standardized condition, but this correlation was better under the improved standardized condition using corrected γ=1 (R(2)=0.9282). Discrepancies in practical HER2 scores were increased in sections with thicknesses <2 and >5 μm. A control system to manage tissue thickness was introduced. CONCLUSIONS: Tissue thickness interferes with the estimation of the IHC intensity of HER2 in both experimental and practical scoring systems. A control system for managing tissue thickness is essential to increase the benefits of IHC as a standardized assay for clinical applications. Lippincott Williams & Wilkins 2021-02 2020-05-12 /pmc/articles/PMC7993914/ /pubmed/32404698 http://dx.doi.org/10.1097/PAI.0000000000000859 Text en Copyright © 2020 The Author(s). Published by Wolters Kluwer Health, Inc. This is an open access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/) (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://creativecommons.org/licenses/by-nc-nd/4.0/ |
spellingShingle | Research Articles Masuda, Shinobu Suzuki, Ryohei Kitano, Yuriko Nishimaki, Haruna Kobayashi, Hiroko Nakanishi, Yoko Yokoi, Hideo Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title | Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title_full | Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title_fullStr | Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title_full_unstemmed | Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title_short | Tissue Thickness Interferes With the Estimation of the Immunohistochemical Intensity: Introduction of a Control System for Managing Tissue Thickness |
title_sort | tissue thickness interferes with the estimation of the immunohistochemical intensity: introduction of a control system for managing tissue thickness |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7993914/ https://www.ncbi.nlm.nih.gov/pubmed/32404698 http://dx.doi.org/10.1097/PAI.0000000000000859 |
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