Epidermal innervation morphometry by immunofluorescence and bright-field microscopy

Maria Nolano, Antonella Biasiotta, Raffaella Lombardi, Vincenzo Provitera, Annamaria Stancanelli, Giuseppe Caporaso, Lucio Santoro, Ingemar S J Merkies, Andrea Truini, Carla Porretta-Serapiglia, Daniele Cazzato, Patrizia Dacci, Dino F. Vitale, Giuseppe Lauria

Research output: Contribution to journalArticlepeer-review


We investigated the agreement between simple indirect immunofluorescence (IF) and bright-field immunohistochemistry (BFI) on free-floating sections for intraepidermal nerve fiber density (IENFD) quantification. Fifty-five healthy subjects and 63 patients with probable small fiber neuropathy (SFN) underwent two adjacent skin biopsies at the distal leg processed by IF and BFI technique. Agreement between IENFD pairs obtained by each method was assessed by Bland-Altman testing. The area under the curve of the receiving operating characteristics (ROC) curves was used to compare the discrimination ability. The diagnostic judgment was based on sex and age-adjusted normative values. IF and BFI showed good correlation (r = 0.81), with a ratio of about 2:1 and a mean difference of 5.5 ± 3.0 IENF per millimeter between paired measures, as demonstrated by linear regression and Bland-Altman test analyses. The square root transformation confirmed a Poisson distribution of the data and a fixed bias between IF and BFI measurements. The ROC curves analysis demonstrated a striking overlap between IF and BFI (0.83 and 0.82; p = 0.72). The diagnosis of SFN disagreed in only 6.7% of cases when the judgment was based on a difference of >1 IENF from 5% cut-off value. IF and BFI showed comparable diagnostic efficiency when referred to appropriate normative reference values.

Original languageEnglish
Pages (from-to)387-391
Number of pages5
JournalJournal of the Peripheral Nervous System
Issue number4
Publication statusPublished - Dec 1 2015


  • immunofluorescence
  • immunohistochemistry
  • intraepidermal nerve fiber
  • painful neuropathy
  • skin biopsy

ASJC Scopus subject areas

  • Clinical Neurology
  • Neuroscience(all)


Dive into the research topics of 'Epidermal innervation morphometry by immunofluorescence and bright-field microscopy'. Together they form a unique fingerprint.

Cite this