Refined Quantitation of Sweat Gland Innervation
Refined Quantitation of Sweat Gland Innervation
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DOI：
10.1093/jnen/nlz015
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发表时间：
2019-05-01
影响因子：
3.2
通讯作者：
Singer, Wolfgang
中科院分区：
文献类型：
--
作者：
Minota, Karla;Schmeichel, Ann M.;Singer, Wolfgang
皮肤活检作为一种评估影响小神经纤维的疾病的工具越来越受欢迎。虽然有关汗腺神经纤维密度(SGNFD)以定量运动神经支配的报道很有希望，但方法学差异很大。虽然传统的体视学被普遍使用，但还没有标准的技术被建立。我们试图开发一种准确和可重复性的技术来定量SGNFD。取自健康人的皮肤穿孔活检组织进行了切割和染色。汗腺(SGS)的图像通过共焦显微镜和广域显微镜获取，并使用去卷积进行优化。用三维(3D)自动化软件重建神经纤维，测量神经纤维长度(NFL)。将NFL除以SG体积得到SGNFD。采用体视学方法对SGNFD进行评估，并进行比较。对10名健康受试者的92个SGs进行了独立观察者的分析。使用共聚焦显微镜，该软件可以可靠地追踪神经纤维。相比之下，神经纤维在广域显微镜下的显示效果较差。与共聚焦图像相比，使用宽视场图像的观察者间可靠性次之(ICC=0.82比ICC=0.98)。三维重建与体视学的相关性较差(ICC=0.38)。新发展的SGNFD定量技术是利用共聚焦显微镜对SG神经支配进行三维重建，可靠地追踪神经纤维，显示出出色的重复性，几乎完全无偏，优于传统的体视学方法。
Skin biopsies have gained increasing popularity as a tool to evaluate disorders affecting small nerve fibers. While reports on sweat gland nerve fiber density (SGNFD) to quantitate sudomotor innervation have been promising, methodologies vary significantly. Although conventional stereology is commonly used, no standard technique has been established. We sought to develop an accurate and reproducible technique to quantify SGNFD. Skin punch biopsies from healthy individuals were cut and stained. Images of sweat glands (SGs) were acquired using confocal and widefield microscopes, and optimized using deconvolution. Nerve fibers were reconstructed and nerve fiber length (NFL) was quantified using three-dimensional (3D) automated software. SGNFD was obtained by dividing NFL by SG volume. SGNFD was also assessed using stereology for comparison. Ninety-two SGs from 10 healthy subjects were analyzed by independent observers. Using confocal microscopy, the software reliably traced nerve fibers. In contrast, rendering of nerve fibers was inferior using widefield microscopy. Interobserver reliability was suboptimal using widefield images compared to confocal (ICC = 0.82 vs ICC = 0.98). Correlation between 3D-reconstruction and stereology was poor (ICC = 0.38). The newly developed technique of SGNFD quantitation using 3D reconstruction of SG innervation with confocal microscopy reliably traces nerve fibers, shows outstanding reproducibility, is almost completely unbiased, and superior to conventional stereology methods.