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Assessing fractional hair cell survival in archival human temporal bones

机译:评估档案人类骨骼的分数毛细胞生存

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Objectives/Hypothesis Histopathological analysis of hair cell survival in human temporal bone sections has historically been binarized such that each hair cell row is rated as either present or absent, thereby greatly underestimating the amount of hair cell loss. Here, we describe and validate a technique to reliably assess fractional hair cell survival in archival sections stained with hematoxylin and eosin (H&E) using high‐resolution light microscopy and optical sectioning. Study Design Technique validation. Methods Hair cell counts in archival temporal bone slide sets were performed by several observers using either differential interference contrast (DIC) or confocal microscopy of the endogenous eosin fluorescence in hair cells. As a further cross‐check, additional decelloidinized sections were immunostained with hair cell markers myosin VI and VIIa. Results Cuticular plates and stereocilia bundles are routinely resolvable in DIC imaging of archival H&E‐stained human material using standard research‐grade microscopes, allowing highly accurate counts of fractional hair cell survival that are reproducible across observer and can be verified by confocal microscopy. Conclusions Reanalysis of cases from the classic temporal bone literature on presbycusis suggests that, contrary to prior reports, differences in audiometric patterns may be well explained by the patterns of hair cell loss. Level of Evidence NA Laryngoscope , 130:487–495, 2020
机译:目标/在人颞骨部毛细胞存活的假说组织病理学分析历来被二值化,使得每个毛细胞行被评为存在或不存在,从而大大低估了毛细胞损失的量。这里,我们描述和验证的技术可靠地评估在用苏木精和伊红染色切片档案分数毛细胞存活;利用高分辨率光学显微镜和光学切片(H&安培E)。研究设计技术验证。在档案颞骨滑动套方法毛发细胞计数通过使用微分干涉对比(DIC)或毛细胞内源性荧光曙红的共聚焦显微镜几个观察员进行。作为进一步的交叉检验,额外decelloidinized切片与毛细胞标志物肌凝蛋白VI和VIIa的免疫染色。结果角质层板和纤毛束是在档案H&安培的DIC成像常规分辨;使用标准的研究级显微镜,允许分数毛细胞的存活是跨越观察者可再现的,并且可以通过共聚焦显微镜进行验证的高度准确的计数E-染色的人材料。从上老年性耳聋的经典颞骨文献箱子结论再分析表明相反,先前的报告,在听力模式的差异可以是深受毛细胞损失的模式进行说明。证据NA喉镜的水平,130:487-495 2020年

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