title: Intraepidermal nerve fiber analysis using immunofluorescence with and without confocal microscopy.
abstract: BACKGROUND
  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.
authors.affiliation: "S. Maugeri" Foundation IRCCS, Center of Telese Terme, Via Bagni Vecchi, 1-82037 Telese Terme, Benevento, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Provitera
authors.forename: Maria
authors.initials: M
authors.lastname: Nolano
authors.forename: Annamaria
authors.initials: A
authors.lastname: Stancanelli
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Caporaso
authors.forename: Dino F
authors.initials: DF
authors.lastname: Vitale
authors.forename: Lucio
authors.initials: L
authors.lastname: Santoro
mesh_terms: D001706:Biopsy / Q000379:methods; D005455:Fluorescent Antibody Technique; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods; D018613:Microscopy, Confocal; D009412:Nerve Fibers; D012867:Skin / Q000294:innervation*
keywords: confocal microscopy; immunofluorescence; intraepidermal nerve fibers; morphometry; skin biopsy