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Related Experiment Video

Updated: Apr 5, 2026

Three-dimensional Imaging and Analysis of Mitochondria within Human Intraepidermal Nerve Fibers
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Epidermal innervation morphometry by immunofluorescence and bright-field microscopy.

Maria Nolano1, Antonella Biasiotta2, Raffaella Lombardi3

  • 1"Salvatore Maugeri" Foundation, Institute of Telese Terme (BN), Telese Terme (BN), Italy.

Journal of the Peripheral Nervous System : JPNS
|August 27, 2015
PubMed
Summary

Indirect immunofluorescence (IF) and bright-field immunohistochemistry (BFI) show comparable diagnostic efficiency for intraepidermal nerve fiber density (IENFD) in small fiber neuropathy (SFN) diagnosis. Both methods provide reliable results for IENFD quantification.

Keywords:
immunofluorescenceimmunohistochemistryintraepidermal nerve fiberpainful neuropathyskin biopsy

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Area of Science:

  • Neurology
  • Histopathology

Background:

  • Intraepidermal nerve fiber density (IENFD) is crucial for diagnosing small fiber neuropathy (SFN).
  • Standardizing IENFD quantification methods is essential for accurate diagnosis.

Purpose of the Study:

  • To compare the agreement and diagnostic efficiency of indirect immunofluorescence (IF) and bright-field immunohistochemistry (BFI) for IENFD quantification.
  • To evaluate the clinical utility of these methods in diagnosing SFN.

Main Methods:

  • Adjacent skin biopsies from 55 healthy subjects and 63 SFN patients were processed using IF and BFI.
  • Agreement was assessed using Bland-Altman testing and linear regression.
  • Diagnostic discrimination was evaluated using receiver operating characteristic (ROC) curves.

Main Results:

  • IF and BFI demonstrated good correlation (r=0.81) with a mean difference of 5.5 ± 3.0 IENF/mm.
  • Both methods showed similar diagnostic efficiency, with ROC curve areas of 0.83 (IF) and 0.82 (BFI).
  • SFN diagnosis disagreed in only 6.7% of cases when using a 5% cut-off difference.

Conclusions:

  • IF and BFI are comparable and reliable methods for IENFD quantification.
  • Both techniques exhibit similar diagnostic performance for SFN when compared to normative values.
  • These findings support the use of either IF or BFI for SFN diagnosis.