Intraepidermal nerve fiber analysis using immunofluorescence with and without confocal microscopy

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Introduction: Intraepidermal nerve fiber (IENF) linear density is a reliable tool to detect small-fiber neuropathies. In immunofluorescence, the IENF count is performed typically by computer-assisted nerve tracing on confocal images. Alternatively, the count can be performed directly through the oculars of a standard epifluorescence microscope. We specifically compared measures obtained using the 2 methods. Methods: We compared measures of IENF density in the same 50 skin samples using computer-assisted image analysis and direct count. Results: There was excellent agreement between the 2 methods. Linear regression showed a slope between paired measures virtually equal to 1 (β=0.99). Bland-Altman analysis showed a mean difference (offset) between the measures of 0.46±0.91 fibers/mm. Conclusions: Direct observation with epifluorescence microscopy proved as reliable as the more time-consuming 3-dimensional computer-assisted analysis of confocal digital images for determining IENF density.

Original languageEnglish
Pages (from-to)501-504
Number of pages4
JournalMuscle and Nerve
Issue number4
Publication statusPublished - Apr 1 2015


  • Confocal microscopy
  • Immunofluorescence
  • Intraepidermal nerve fibers
  • Morphometry
  • Skin biopsy

ASJC Scopus subject areas

  • Clinical Neurology
  • Cellular and Molecular Neuroscience
  • Physiology (medical)
  • Physiology
  • Medicine(all)


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