Utilizing image analysis by optical density to evaluate changes in hematoxylin and eosin staining quality after reagent overuse

Due to the prevalence of hematoxylin and eosin (H&E) staining in routine histological preparations, understanding the factors that impact stain color characteristics is vital to attain consistently high-quality stains. In the last decade, increased use of digital pathology and image analysis (specifically by optical density [OD] measures) has provided new ways of assessing staining precision. This paper combines data from two studies that tracked H&E staining quality in both nuclear and cytoplasmic components of 12 tissues (11 human and 1 porcine) by OD after overuse of H&E staining reagents from 5 vendors. Both studies showed a decrease in eosin stain intensity by OD and visual inspection (by a histologist) with reagent overuse. This trend varied in degree by tissue type and reagent vendor. Nonetheless, staining quality for H&E staining from all vendors and for all organs remained acceptable (but not always optimal) for microscopic evaluation by the College of American Pathologists (CAP) and National Society for Histotechnology (NSH) staining criteria when stained sections were reviewed by a board-certified veterinary pathologist.

鉴于苏木精-伊红（H&E）染色在常规组织学制片中的广泛应用，明确影响染色色彩特征的相关因素，对于获得稳定一致的高质量染色结果至关重要。近十年来，数字病理学（digital pathology）与图像分析（image analysis）技术的应用日益增多，尤其是通过光密度（optical density, OD）量化手段，为评估染色精准度提供了全新途径。本研究整合了两项研究的数据：两项研究均通过光密度（OD）检测，对5家供应商提供的H&E染色试剂过度使用后，12种组织（11种人类组织与1种猪源组织）的细胞核与细胞质组分的H&E染色质量进行了追踪评估。两项研究均显示，随着染色试剂过度使用，通过OD检测与组织病理学家目视评估均能发现伊红染色强度出现下降。该下降趋势的严重程度因组织类型与试剂供应商的不同而存在差异。尽管如此，当由认证合格的兽医病理学家对染色切片进行阅片时，按照美国病理学家协会（College of American Pathologists, CAP）与美国组织技术师协会（National Society for Histotechnology, NSH）制定的染色标准进行镜检评估，所有供应商的试剂对所有受试组织的H&E染色质量仍处于可接受水平（但并非始终处于最佳状态）。