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用于无标记成像和定量检测表皮内神经纤维的不连续三次谐波产生显微镜。

Discontinuity third harmonic generation microscopy for label-free imaging and quantification of intraepidermal nerve fibers.

机构信息

Department of Electrical Engineering and Graduate Institute of Photonics and Optoelectronics, National Taiwan University, Taipei 10617, Taiwan.

Department of Neurology, National Taiwan University Hospital, and National Taiwan University College of Medicine, Taipei 100225, Taiwan.

出版信息

Cell Rep Methods. 2024 Mar 25;4(3):100735. doi: 10.1016/j.crmeth.2024.100735. Epub 2024 Mar 18.

DOI:10.1016/j.crmeth.2024.100735
PMID:38503290
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10985268/
Abstract

Label-free imaging methodologies for nerve fibers rely on spatial signal continuity to identify fibers and fail to image free intraepidermal nerve endings (FINEs). Here, we present an imaging methodology-called discontinuity third harmonic generation (THG) microscopy (dTHGM)-that detects three-dimensional discontinuities in THG signals as the contrast. We describe the mechanism and design of dTHGM and apply it to reveal the bead-string characteristics of unmyelinated FINEs. We confirmed the label-free capability of dTHGM through a comparison study with the PGP9.5 immunohistochemical staining slides and a longitudinal spared nerve injury study. An intraepidermal nerve fiber (IENF) index based on a discontinuous-dot-connecting algorithm was developed to facilitate clinical applications of dTHGM. A preliminary clinical study confirmed that the IENF index was highly correlated with skin-biopsy-based IENF density (Pearson's correlation coefficient R = 0.98) and could achieve differential identification of small-fiber neuropathy (p = 0.0102) in patients with diabetic peripheral neuropathy.

摘要

无标记神经纤维成像方法依赖于空间信号连续性来识别纤维，无法对游离表皮神经末梢 (FINEs) 成像。在这里，我们提出了一种成像方法——不连续的三次谐波产生 (THG) 显微镜 (dTHGM)——它检测 THG 信号中的三维不连续性作为对比度。我们描述了 dTHGM 的机制和设计，并将其应用于揭示无髓鞘 FINEs 的珠串特征。我们通过与 PGP9.5 免疫组织化学染色载玻片和纵向神经损伤保留研究的比较研究证实了 dTHGM 的无标记能力。开发了一种基于不连续点连接算法的表皮内神经纤维 (IENF) 指数，以促进 dTHGM 的临床应用。一项初步的临床研究证实，IENF 指数与皮肤活检的 IENF 密度高度相关（Pearson 相关系数 R = 0.98），并且可以区分糖尿病周围神经病变患者的小纤维神经病（p = 0.0102）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/be9fd305df74/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/b61ce207e689/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/211ef530b9bc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/bda88a95eeea/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/59c9fbbfa8d5/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/96b8a7d9570d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/4cbb4761b981/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/b3512cc1b7ec/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/be9fd305df74/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/b61ce207e689/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/211ef530b9bc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/bda88a95eeea/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/59c9fbbfa8d5/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/96b8a7d9570d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/4cbb4761b981/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/b3512cc1b7ec/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4e/10985268/be9fd305df74/gr7.jpg

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