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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2025-05-01
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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 |
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| 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. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-05-01 |
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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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