Intraepidermal nerve fiber analysis using immunofluorescence with and without confocal microscopy
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Updated: Apr 26, 2026
Three-dimensional Imaging and Analysis of Mitochondria within Human Intraepidermal Nerve Fibers
Published on: September 29, 2017
Vincenzo Provitera1, Maria Nolano, Annamaria Stancanelli
1"S. Maugeri" Foundation IRCCS, Center of Telese Terme, Via Bagni Vecchi, 1-82037 Telese Terme, Benevento, Italy.
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.
