QuPath Automated Analysis of Optic Nerve Degeneration in Brown Norway Rats.
QuPath Automated Analysis of Optic Nerve Degeneration in Brown Norway Rats.
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QuPath 自动分析棕色挪威大鼠视神经变性。
DOI：
10.1167/tvst.9.3.22
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发表时间：
2020
影响因子：
3
通讯作者：
Bollinger,KathrynE
中科院分区：
文献类型：
--
作者：
Mysona,BarbaraA;Segar,Sharmila;Hernandez,Cecilia;Kim,Christian;Zhao,Jing;Mysona,David;Bollinger,KathrynE
目的：QuPath 开源数字分析软件的新应用用于对大鼠进行性视神经 (ON) 变性进行深入的形态学分析。方法：QuPath 软件适用于评估甲苯胺蓝染色的棕色挪威大鼠 ON 光学显微照片中的轴突和神经胶质细胞形态。 QuPath 轴突数量、密度、尺寸分布和神经胶质细胞区域是从测试图像和按损伤等级划分的 ON 横截面获得的。将 QuPath 结果与手动计数、AxonJ 和电子显微镜轴突估计值进行比较。结果：QuPath 衍生的轴突数量、密度和直径随着 ON 损伤的增加而减少。轴突密度与测试图像中的神经胶质细胞区域呈负相关（R 2 = 0.759；P < 0.0001；N = 40）和 ON 横截面（R 2 = 0.803；P < 0.0004；N = 10）。尽管轴突损失发生在大多数轴突直径上，但大轴突更容易退化。例外是肿胀的轴突> 2 µm，在中度但不严重受损的图像中肿胀轴突增加。 QuPath 轴突计数与测试图像的手动计数密切相关（R 2= 0.956；P< 0.0001）。 QuPath 在测试图像和 ON 轴突总数方面优于 AxonJ。与电子显微镜分析相比，QuPath 低估了 ON 轴突；然而，这些方法之间的相关性很强（R 2 = 0.797；P < 0.001；N = 10）。结论：QuPath 分析可靠地识别了退化 ON 中发生的轴突丢失、轴突形态变化和神经胶质扩张。翻译相关性：QuPath 是快速、自动化分析健康和退化 ON 的有价值的工具。新青光眼治疗方法的可重复临床前研究取决于对视神经病理学的公正深入分析。这是由 QuPath 方法提供的。
Purpose: A novel application of QuPath open-source digital analysis software is used to provide in-depth morphological analysis of progressive optic nerve (ON) degeneration in rats.Methods: QuPath software was adapted to assess axon and gliotic morphology in toluidine blue-stained, Brown Norway rat ON light micrographs. QuPath axon numbers, density, size distributions, and gliotic areas were obtained from test images and ON cross-sections separated by damage grade. QuPath results were compared with manual counting, AxonJ, and electron microscopy axon estimates.Results: QuPath-derived axon number, density, and diameter decreased with increasing ON damage. Axon density negatively correlated with gliotic areas in test images (R 2= 0.759; P< 0.0001; N= 40) and in ON cross-sections (R 2= 0.803; P< 0.0004; N= 10). Although axon losses occurred across most axon diameters, large axons were more susceptible to degeneration. The exception was swollen axons> 2 µm, which increased in moderately but not severely damaged images. QuPath axon counts correlated strongly with manual counts of test images (R 2= 0.956; P< 0.0001). QuPath outperformed AxonJ on test images and total ON axon counts. Compared to electron microscopy analysis, QuPath undercounted ON axons; however, correlation between the methods was robust (R 2= 0.797; P< 0.001; N= 10).Conclusions: QuPath analysis reliably identified axon loss, axon morphology changes, and gliotic expansion that occurred in degenerating ONs.Translational Relevance: QuPath is a valuable tool for rapid, automated, analysis of healthy and degenerating ONs. Reproducible preclinical studies for new glaucoma treatments depend on unbiased in-depth analysis of ON pathology. This was provided by the QuPath approach.