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用共聚焦显微镜测量新鲜和保存的人耳膜的厚度分布。

Thickness distribution of fresh and preserved human eardrums measured with confocal microscopy.

作者信息

Kuypers Liesbeth C, Decraemer Willem F, Dirckx Joris J J

机构信息

Department of Physics, University of Antwerp, Antwerpen, Belgium.

出版信息

Otol Neurotol. 2006 Feb;27(2):256-64. doi: 10.1097/01.mao.0000187044.73791.92.

Abstract

HYPOTHESIS

In this study, the thickness distribution of the fresh human eardrum was measured and possible thickness changes in successive stages of preservation and preparation were studied.

METHODS

The thickness measurement was performed on axial fluorescence images taken perpendicularly through the membrane with a confocal microscope. The influence of fixation and preservation (in Cialit solution) on the thickness was also investigated. The same eardrum was prepared (decalcified, dehydrated, and stained) for histologic sectioning and the thickness was measured on the sections using conventional light microscopy.

RESULTS

Similar thickness distributions in the measured samples (n = 3) were observed. The pars tensa has a rather constant thickness in a central region curving as a horseshoe upward around the manubrium. The membrane thickens slightly from the inferior to the superior side. The anterior region is thicker than the posterior region. In narrow bands along the manubrium, peripheral rim, and in the region inferior to the umbo, a much larger thickness in comparison with that in the central region was found. Mean thicknesses of approximately 40, 50, and 120 microm were observed in the central region of the studied eardrums, respectively.

CONCLUSION

Whereas the thickness distribution of the human eardrums shows similar features, the absolute thickness seems to vary a lot from one specimen to another. There is no significant difference in thickness of the same membrane in fresh, fixed, or preserved condition. Thus, human eardrums may be safely preserved in fixative for later thickness measurements. The histologic preparation process, however, causes a significant location-dependent shrinkage.

摘要

假设

在本研究中,测量了新鲜人耳膜的厚度分布,并研究了保存和制备连续阶段可能发生的厚度变化。

方法

使用共聚焦显微镜对垂直穿过耳膜拍摄的轴向荧光图像进行厚度测量。还研究了固定和保存(在西力替溶液中)对厚度的影响。将同一张耳膜进行制备(脱钙、脱水和染色)以进行组织切片,并使用传统光学显微镜在切片上测量厚度。

结果

在所测量的样本(n = 3)中观察到相似的厚度分布。紧张部在围绕柄向上弯曲成马蹄形的中央区域具有相当恒定的厚度。耳膜从下侧到上侧略有增厚。前部区域比后部区域厚。在沿着柄、周边边缘以及在鼓膜脐下方的狭窄带中,发现与中央区域相比厚度大得多。在所研究的耳膜中央区域分别观察到平均厚度约为40、50和120微米。

结论

虽然人耳膜的厚度分布显示出相似的特征,但绝对厚度似乎因标本而异。新鲜、固定或保存状态下同一耳膜的厚度没有显著差异。因此,人耳膜可以安全地保存在固定剂中以便以后进行厚度测量。然而,组织学制备过程会导致显著的位置依赖性收缩。

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