Department of Pathology, Osaka University Graduate School of Medicine, Osaka, Japan; Department of Immunopathology, WPI Immunology Frontier Research Center, Osaka University, Osaka, Japan.
Graduate School of Medical Life Science, Yokohama City University, Kanagawa, Japan.
Cell Mol Gastroenterol Hepatol. 2022;14(4):905-924. doi: 10.1016/j.jcmgh.2022.07.001. Epub 2022 Jul 12.
BACKGROUND & AIMS: Tissue-clearing and three-dimensional (3D) imaging techniques aid clinical histopathological evaluation; however, further methodological developments are required before use in clinical practice.
We sought to develop a novel fluorescence staining method based on the classical periodic acid-Schiff stain. We further attempted to develop a 3D imaging system based on this staining method and evaluated whether the system can be used for quantitative 3D pathological evaluation and deep learning-based automatic diagnosis of inflammatory bowel diseases.
We successfully developed a novel periodic acid-FAM hydrazide (PAFhy) staining method for 3D imaging when combined with a tissue-clearing technique (PAFhy-3D). This strategy enabled clear and detailed imaging of the 3D architectures of crypts in human colorectal mucosa. PAFhy-3D imaging also revealed abnormal architectural changes in crypts in ulcerative colitis tissues and identified the distributions of neutrophils in cryptitis and crypt abscesses. PAFhy-3D revealed novel pathological findings including spiral staircase-like crypts specific to inflammatory bowel diseases. Quantitative analysis of crypts based on 3D morphologic changes enabled differential diagnosis of ulcerative colitis, Crohn's disease, and non-inflammatory bowel disease; such discrimination could not be achieved by pathologists. Furthermore, a deep learning-based system using PAFhy-3D images was used to distinguish these diseases The accuracies were excellent (macro-average area under the curve = 0.94; F1 scores = 0.875 for ulcerative colitis, 0.717 for Crohn's disease, and 0.819 for non-inflammatory bowel disease).
PAFhy staining and PAFhy-3D imaging are promising approaches for next-generation experimental and clinical histopathology.
组织透明化和三维（3D）成像技术有助于临床组织病理学评估；然而，在临床实践中应用之前，还需要进一步的方法学发展。
我们试图开发一种基于经典过碘酸-Schiff 染色的新型荧光染色方法。我们进一步尝试开发一种基于这种染色方法的 3D 成像系统，并评估该系统是否可用于定量 3D 病理评估和基于深度学习的炎症性肠病自动诊断。
我们成功开发了一种新型的过碘酸-FAM 酰肼（PAFhy）染色方法，可与组织透明化技术（PAFhy-3D）结合使用。该策略可实现人结直肠黏膜中隐窝 3D 结构的清晰和详细成像。PAFhy-3D 成像还揭示了溃疡性结肠炎组织中隐窝的异常结构改变，并确定了隐窝炎和隐窝脓肿中中性粒细胞的分布。PAFhy-3D 揭示了包括炎症性肠病特有的螺旋梯状隐窝在内的新的病理发现。基于 3D 形态变化的隐窝定量分析可实现溃疡性结肠炎、克罗恩病和非炎症性肠病的鉴别诊断；病理学家无法实现这种区分。此外，使用 PAFhy-3D 图像的基于深度学习的系统用于区分这些疾病，准确率非常高（宏观平均曲线下面积为 0.94；溃疡性结肠炎的 F1 得分为 0.875，克罗恩病为 0.717，非炎症性肠病为 0.819）。
PAFhy 染色和 PAFhy-3D 成像有望成为下一代实验和临床组织病理学的方法。
