Premier Laboratory, LLC, Longmont, CO, USA.
Boulder Statistics, LLC, Steamboat Springs, CO, USA.
J Histotechnol. 2020 Mar;43(1):29-37. doi: 10.1080/01478885.2019.1708611. Epub 2020 Jan 23.
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.
染色质量和可重复性是监测实验室质量保证的重要因素。在过去的十年中，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通过光密度（OD）跟踪染色精度的潜在手段，但也带来了挑战。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。本研究有两个目的。第一个目的是通过 OD 跟踪核和细胞质成分中苏木精和伊红（H&E）染色的精度。第二个目的是确定不同的分析前和分析变量对 OD 结果的影响。具体而言，研究的终点包括质量参数，包括切片厚度、方案操作、过期苏木精对染色精度和随时间变化的染色重现性的影响。我们的结果表明，对 H&E 染色组织切片进行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。这些作者选择了图形而不是完全的统计分析结果，以突出视觉辅助工具在演示 H&E 染色重现性方面的实用性。
