Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer

Accurately determining HER2 status is essential for breast cancer treatment. We developed an AI-integrated in-house application for automated Dual bright-field (BF) in situ hybridization (ISH) analysis on whole slide images (WSIs), although optimal scanning conditions remain unclear. We evaluated sc...

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Main Authors: Nilay Bakoglu Malinowski, Takashi Ohnishi, Emine Cesmecioglu, Dara S. Ross, Tetsuya Tsukamoto, Yukako Yagi
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Bioengineering
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Online Access:https://www.mdpi.com/2306-5354/12/6/569
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author Nilay Bakoglu Malinowski
Takashi Ohnishi
Emine Cesmecioglu
Dara S. Ross
Tetsuya Tsukamoto
Yukako Yagi
author_facet Nilay Bakoglu Malinowski
Takashi Ohnishi
Emine Cesmecioglu
Dara S. Ross
Tetsuya Tsukamoto
Yukako Yagi
author_sort Nilay Bakoglu Malinowski
collection DOAJ
description Accurately determining HER2 status is essential for breast cancer treatment. We developed an AI-integrated in-house application for automated Dual bright-field (BF) in situ hybridization (ISH) analysis on whole slide images (WSIs), although optimal scanning conditions remain unclear. We evaluated scanners and optimized scanning protocols for clinical application. Ten de-identified invasive breast carcinoma cases, with HER2 immunohistochemistry and FISH results, were analyzed using three scanners and six scanning protocols. WSIs scanned by Scanner ‘A’ have 0.12 µm/pixel with 0.95 NA (A1) and 1.2 NA (A2); Scanner ‘B’ have 0.08 µm/pixel (B1); 0.17 µm/pixel (B2); and 0.17 µm/pixel with extended focus (1.4 µm step size and three layers) (B3); Scanner ‘C’ has 0.26 µm/pixel (C1) resolution. Results showed scanning protocols A1, A2, B2, and B3 yielded HER2 gene amplification status and ASCO/CAP ISH group results consistent with manual FISH as the ground truth. However, protocol C demonstrated poor concordance due to nuclei detection failure in six cases. The AI-integrated application achieved the best performance using scanning protocols with optimized resolutions of 0.12 µm/pixel and 0.17 µm/pixel with extended focus. This study highlights the importance of scanner selection in AI-based HER2 assessment and demonstrates that optimized scanning parameters enhance the accuracy and reliability of automated Dual BF ISH analysis.
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spelling doaj-art-1f26c48f7a334ddbac5fcd15717ef72d2025-08-20T03:27:26ZengMDPI AGBioengineering2306-53542025-05-0112656910.3390/bioengineering12060569Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast CancerNilay Bakoglu Malinowski0Takashi Ohnishi1Emine Cesmecioglu2Dara S. Ross3Tetsuya Tsukamoto4Yukako Yagi5Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USADepartment of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USADepartment of Pathology, Marmara University Research and Education Hospital, 34899 Istanbul, TurkeyDepartment of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USADepartment of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USADepartment of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USAAccurately determining HER2 status is essential for breast cancer treatment. We developed an AI-integrated in-house application for automated Dual bright-field (BF) in situ hybridization (ISH) analysis on whole slide images (WSIs), although optimal scanning conditions remain unclear. We evaluated scanners and optimized scanning protocols for clinical application. Ten de-identified invasive breast carcinoma cases, with HER2 immunohistochemistry and FISH results, were analyzed using three scanners and six scanning protocols. WSIs scanned by Scanner ‘A’ have 0.12 µm/pixel with 0.95 NA (A1) and 1.2 NA (A2); Scanner ‘B’ have 0.08 µm/pixel (B1); 0.17 µm/pixel (B2); and 0.17 µm/pixel with extended focus (1.4 µm step size and three layers) (B3); Scanner ‘C’ has 0.26 µm/pixel (C1) resolution. Results showed scanning protocols A1, A2, B2, and B3 yielded HER2 gene amplification status and ASCO/CAP ISH group results consistent with manual FISH as the ground truth. However, protocol C demonstrated poor concordance due to nuclei detection failure in six cases. The AI-integrated application achieved the best performance using scanning protocols with optimized resolutions of 0.12 µm/pixel and 0.17 µm/pixel with extended focus. This study highlights the importance of scanner selection in AI-based HER2 assessment and demonstrates that optimized scanning parameters enhance the accuracy and reliability of automated Dual BF ISH analysis.https://www.mdpi.com/2306-5354/12/6/569artificial intelligencedual bright-field in-situ hybridizationbreast carcinomawhole slide imagescanning protocol
spellingShingle Nilay Bakoglu Malinowski
Takashi Ohnishi
Emine Cesmecioglu
Dara S. Ross
Tetsuya Tsukamoto
Yukako Yagi
Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
Bioengineering
artificial intelligence
dual bright-field in-situ hybridization
breast carcinoma
whole slide image
scanning protocol
title Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
title_full Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
title_fullStr Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
title_full_unstemmed Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
title_short Optimization of Scanning Protocol for AI-Integrated Assessment of HER2 Dual Bright-Field In-Situ Hybridization Application in Breast Cancer
title_sort optimization of scanning protocol for ai integrated assessment of her2 dual bright field in situ hybridization application in breast cancer
topic artificial intelligence
dual bright-field in-situ hybridization
breast carcinoma
whole slide image
scanning protocol
url https://www.mdpi.com/2306-5354/12/6/569
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