Comparative quantitative assessment of the human corneal sub-basal nerve plexus by in vivo confocal microscopy and histological staining
Comparative quantitative assessment of the human corneal sub-basal nerve plexus by in vivo confocal microscopy and histological staining
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DOI：
10.1038/eye.2016.220
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发表时间：
2017-03-01
期刊：
影响因子：
3.9
通讯作者：
Guthoff, R. F.
中科院分区：
文献类型：
--
作者：
Kowtharapu, B. S.;Winter, K.;Guthoff, R. F.
目的比较两种不同方法测量人角膜基底下神经丛（sub-basal nerve plexus，SBP）的定量数据：体内共聚焦显微镜（IVCM），方法将来自健康受试者的大规模IVCM镶嵌图像中的SBP的七个参数与用a-III-Doppler染色的离体供体角膜中的相同参数进行比较。微管蛋白免疫组织化学角膜神经纤维长度（CNFL）、角膜神经纤维密度（CNFD）、角膜神经分支密度（CNBD）、平均加权角膜神经纤维弯曲度（CNFTo）、角膜神经连接点（CNCP）、平均角膜神经单纤维长度（CNSFL）和平均加权角膜神经纤维厚度（CNFTh）使用专用的，结果我们的实验表明CNFL的值显著高于其他算法（50.2 vs 21.4 mm/mm（2）），CNFD（1358.8 vs 277.3神经纤维/mm（2）），CNBD（847.6 vs 163.5分支/mm（2）），CNFTo（0.095 vs 0.081 μ m（-1））和CNCP（49.4 vs 21.6连接/mm（2））。相比之下，CNSFL值高于IVCM图像比组织学标本（32.1与74.1 μ m）。CNFTh（2.22 μ m与2.20 μ m）之间没有显着差异，观察到两个groups.Conclusions本研究的结果表明，IVCM具有固有的较低的分辨率与离体免疫组化染色的角膜SBP相比，这种限制导致系统低估的几个SBP参数。尽管有这个缺点，IVCM是一个重要的临床工具，在体内表征，定量临床成像，并评价人角膜SBP。
Purpose This study was designed to compare and contrast quantitative data of the human corneal sub-basal nerve plexus (SBP) evaluated by two different methods: in vivo confocal microscopy (IVCM), and immunohistochemical staining of ex vivo donor corneas.Methods Seven parameters of the SBP in large-scale IVCM mosaicking images from healthy subjects were compared with the identical parameters in ex vivo donor corneas stained by a-III-tubulin immunohistochemistry. Corneal nerve fiber length (CNFL), corneal nerve fiber density (CNFD), corneal nerve branch density (CNBD), average weighted corneal nerve fiber tortuosity (CNFTo), corneal nerve connection points (CNCP), average corneal nerve single-fiber length (CNSFL), and average weighted corneal nerve fiber thickness (CNFTh) were calculated using a dedicated, published algorithm and compared.Results Our experiments showed significantly higher values for CNFL (50.2 vs 21.4 mm/mm(2)), CNFD (1358.8 vs 277.3 nerve fibers/mm(2)), CNBD (847.6 vs 163.5 branches/mm(2)), CNFTo (0.095 vs 0.081 mu m(-1)), and CNCP (49.4 vs 21.6 connections/mm(2)) in histologically staining specimens compared with IVCM images. In contrast, CNSFL values were higher in IVCM images than in histological specimens (32.1 vs 74.1 mu m). No significant difference was observed in CNFTh (2.22 vs 2.20 mu m) between the two groups.Conclusions The results of this study have shown that IVCM has an inherently lower resolution compared with ex vivo immunohistochemical staining of the corneal SBP and that this limitation leads to a systematic underestimation of several SBP parameters. Despite this shortcoming, IVCM is a vital clinical tool for in vivo characterization, quantitative clinical imaging, and evaluation of the human corneal SBP.