Tissue-Clearing Technique and Cutaneous Nerve Biopsies: Quantification of the Intraepidermal Nerve-Fiber Density Using Active Clarity Technique-Pressure Related Efficient and Stable Transfer of Macromolecules Into Organs
10.3988/jcn.2019.15.4.537
- Author:
Dai Hyun KIM
1
;
Se Jeong LEE
;
Eunsoo LEE
;
Ji Hyuck HONG
;
Soo Hong SEO
;
Hyo Hyun AHN
;
Byung Jo KIM
;
Woong SUN
;
Im Joo RHYU
Author Information
1. Department of Anatomy, Korea University College of Medicine, Seoul, Korea. irhyu@korea.ac.kr
- Publication Type:Original Article
- Keywords:
cutaneous nerve biopsy;
peripheral neuropathy;
tissue-clearing and labeling technique;
ACT-PRESTO;
intraepidermal nerve-fiber density
- MeSH:
Biopsy;
Fluorescent Antibody Technique, Indirect;
Humans;
Imaging, Three-Dimensional;
Immunohistochemistry;
Methods;
Nerve Fibers;
Neuralgia, Postherpetic;
Peripheral Nerves;
Peripheral Nervous System Diseases;
Prospective Studies
- From:Journal of Clinical Neurology
2019;15(4):537-544
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND AND PURPOSE: Cutaneous nerve biopsies based on two-dimensional analysis have been regarded as a creditable assessment tool for diagnosing peripheral neuropathies. However, advancements in methodological imaging are required for the analysis of intact structures of peripheral nerve fibers. A tissue-clearing and labeling technique facilitates three-dimensional imaging of internal structures in unsectioned, whole biological tissues without excessive time or labor costs. We sought to establish whether a tissue-clearing and labeling technique could be used for the diagnostic evaluation of peripheral neuropathies. METHODS: Five healthy individuals and four patients with small-fiber neuropathy (SFN) and postherpetic neuralgia (PHN) were prospectively enrolled. The conventional methods of indirect immunofluorescence (IF) and bright-field immunohistochemistry (IHC) were adopted in addition to the tissue-clearing and labeling method called active clarity technique-pressure related efficient and stable transfer of macromolecules into organs (ACT-PRESTO) to quantify the intraepidermal nerve-fiber density (IENFD). RESULTS: The mean IENFD values obtained by IF, bright-field IHC, and ACT-PRESTO in the healthy control group were 6.54, 6.44, and 90.19 fibers/mm², respectively; the corresponding values in the patients with SFN were 1.99, 2.32, and 48.12 fibers/mm², respectively, and 3.06, 2.87, and 47.21 fibers/mm², respectively, in the patients with PHN. CONCLUSIONS: This study has shown that a tissue-clearing method provided not only rapid and highly reproducible three-dimensional images of cutaneous nerve fibers but also yielded reliable quantitative IENFD data. Quantification of the IENFD using a tissue-clearing and labeling technique is a promising way to improve conventional cutaneous nerve biopsies.
