ABSTRACT Staining quality and reproducibility are essential factors to monitor laboratory quality assurance. In the last decade, there has been an increase in the use of digital pathology and image analysis. While the adoption of these tools provides a potential means to track staining precision by optical density (OD), it also presents challenges. Results from image analysis are more sensitive to variations in staining than microscopic evaluation by a pathologist. There are two goals with this study. The first was to track the precision of hematoxylin and eosin (H&E) staining, in both nuclear and cytoplasmic components by OD. The second was to determine the impact of different pre-analytical and analytical variables on the OD results. Specifically, the endpoints investigated were quality parameters including impacts of section thickness, protocol manipulation, expired hematoxylin on staining precision and reproducibility of staining over time. Our results show that image analysis of H&E-stained tissue sections is a viable tool for assessing and verifying staining quality. We also show that OD analysis results for H&E-stained sections are affected by changing pre-analytical and/or reagent variables. These authors chose a graphical rather than fully statistical analysis of the results to highlight the utility of visual aids in demonstrating H&E staining reproducibility.

中文翻译：

---

用于评价苏木精和伊红染色精度的基于光密度的图像分析方法

摘要 染色质量和重现性是监测实验室质量保证的重要因素。在过去十年中，数字病理学和图像分析的使用有所增加。虽然这些工具的采用提供了一种通过光密度 (OD) 跟踪染色精度的潜在方法，但它也带来了挑战。与病理学家的显微镜评估相比，图像分析的结果对染色的变化更敏感。这项研究有两个目标。第一个是通过 OD 在核和细胞质组分中跟踪苏木精和伊红 (H&E) 染色的精确度。第二个是确定不同的预分析和分析变量对 OD 结果的影响。具体而言，研究的终点是质量参数，包括切片厚度的影响、协议操作，过期苏木精对染色精度和染色重现性随时间的影响。我们的结果表明，H&E 染色组织切片的图像分析是评估和验证染色质量的可行工具。我们还表明，H&E 染色切片的 OD 分析结果受分析前和/或试剂变量变化的影响。这些作者选择对结果进行图形分析而不是完全统计分析，以强调视觉辅助工具在证明 H&E 染色再现性方面的效用。E 染色切片受分析前和/或试剂变量变化的影响。这些作者选择对结果进行图形分析而不是完全统计分析，以强调视觉辅助工具在证明 H&E 染色再现性方面的效用。E 染色切片受分析前和/或试剂变量变化的影响。这些作者选择对结果进行图形分析而不是完全统计分析，以强调视觉辅助工具在证明 H&E 染色再现性方面的效用。