Assessing fractional hair cell survival in archival human temporal bones.
Assessing fractional hair cell survival in archival human temporal bones.
复制标题
评估档案人类颞骨中毛细胞的存活率。
DOI：
10.1002/lary.27991
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Liberman,MCharles
中科院分区：
文献类型：
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作者：
Wu,Pei-Zhe;Wen,Wei-Ping;O'Malley,JenniferT;Liberman,MCharles
目的/假设人类颞骨切片中毛细胞存活的组织病理学分析在历史上被二值化，使得每一行毛细胞被评为存在或不存在，从而极大地低估了毛细胞的损失量。在这里，我们描述并验证了一种使用高分辨率光学显微镜和光学切片来可靠地评估苏木精-伊红(H&E)染色的档案切片中毛细胞存活的技术。研究设计技术验证。方法由几个观察者使用差示干涉对比(DIC)或共聚焦显微镜进行毛细胞内源性曙红荧光的计数。结果在使用标准研究级显微镜对H&E染色的档案材料进行DIC成像时，角质板和立体纤毛束通常是可分辨的，允许高精度的毛细胞存活分数的计数，这些计数可在观察者中重现，并可通过共焦显微镜进行验证。结论对经典颞骨文献中关于老年性耳聋的病例的再分析表明，与先前的报道相反，听力模式的差异可以由毛细胞丢失的模式很好地解释。
Objectives/HypothesisHistopathological 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 DesignTechnique validation.MethodsHair 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.ResultsCuticular 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.ConclusionsReanalysis 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 EvidenceNALaryngoscope, 130:487–495, 2020