Supplemental Digital Content is available in the text. With immunohistochemical (IHC) staining increasingly being used to guide clinical decisions, variability in staining quality and reproducibility are becoming essential factors in generating diagnoses using IHC tissue preparations. The current study tested a method to track and quantify the interrun, intrarun, and intersite variability of IHC staining intensity. Our hypothesis was that staining precision between laboratory sites, staining runs, and individual slides may be verified quantitatively, efficiently and effectively utilizing algorithm-based, automated image analysis. To investigate this premise, we tested the consistency of IHC staining in 40 routinely processed (formalin-fixed, paraffin-embedded) human tissues using 10 common antibiomarker antibodies on 2 Dako Omnis instruments at 2 locations (Carpinteria, CA: 30 m above sea level and Longmont, CO: 1500 m above sea level) programmed with identical, default settings and sample pretreatments. Digital images of IHC-labeled sections produced by a whole slide scanner were analyzed by a simple commercially available algorithm and compared with a board-certified veterinary pathologist’s semiquantitative scoring of staining intensity. The image analysis output correlated well with pathology scores but had increased sensitivity for discriminating subtle variations and providing reproducible digital quantification across sites as well as within and among staining runs at the same site. Taken together, our data indicate that digital image analysis offers an objective and quantifiable means of verifying IHC staining parameters as a part of laboratory quality assurance systems.

中文翻译：

---

用于定量和确定免疫组织化学染色重现性的图像分析解决方案

文本中提供了补充数字内容。随着免疫组织化学 (IHC) 染色越来越多地用于指导临床决策，染色质量和重现性的可变性正在成为使用 IHC 组织制剂进行诊断的重要因素。目前的研究测试了一种跟踪和量化 IHC 染色强度的运行间、运行内和位点间变异性的方法。我们的假设是，实验室地点、染色运行和单个载玻片之间的染色精度可以通过基于算法的自动图像分析进行定量、高效和有效的验证。为了研究这个前提，我们测试了 40 种常规处理的 IHC 染色的一致性（福尔马林固定，石蜡包埋）人体组织，在 2 个位置（加利福尼亚州卡宾特里亚：海拔 30 米和科罗拉多州朗蒙特：海拔 1500 米）的 2 台 Dako Omnis 仪器上使用 10 种常见的抗生物标志物抗体，使用相同的默认设置和样品预处理进行编程。通过简单的商用算法分析由整个玻片扫描仪产生的 IHC 标记切片的数字图像，并与获得委员会认证的兽医病理学家的染色强度半定量评分进行比较。图像分析输出与病理学评分良好相关，但在区分细微变化和提供跨位点以及同一位点染色运行内部和之间的可重复数字量化方面具有更高的灵敏度。综合起来，