TY - JOUR
T1 - Tissue-clearing technique and cutaneous nerve biopsies
T2 - Quantification of the intraepidermal nerve-fiber density using active clarity technique-pressure related efficient and stable transfer of macromolecules into organs
AU - Kim, Dai Hyun
AU - Lee, Se Jeong
AU - Lee, Eunsoo
AU - Hong, Ji Hyuck
AU - Seo, Soo Hong
AU - Ahn, Hyo Hyun
AU - Kim, Byung Jo
AU - Sun, Woong
AU - Rhyu, Im Joo
N1 - Funding Information:
This study was supported by the Brain Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Science, ICT, & Future Planning (NRF-2015M3C7A1028790) and Korea University Grant K1710481.
Publisher Copyright:
© 2019 Korean Neurological Association.
PY - 2019/10
Y1 - 2019/10
N2 - 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.
AB - 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.
KW - ACT-PRESTO
KW - Intraepidermal nerve-fiber density
KW - Peripheral neuropathy
KW - Tissue-clearing and labeling technique
KW - Cutaneous nerve biopsy
UR - http://www.scopus.com/inward/record.url?scp=85073514152&partnerID=8YFLogxK
U2 - 10.3988/jcn.2019.15.4.537
DO - 10.3988/jcn.2019.15.4.537
M3 - Article
AN - SCOPUS:85073514152
SN - 1738-6586
VL - 15
SP - 537
EP - 544
JO - Journal of Clinical Neurology (Korea)
JF - Journal of Clinical Neurology (Korea)
IS - 4
ER -