Abstract
We developed a time-efficient semi-automated axon quantification method using freeware in human cranial nerve sections stained with paraphenylenediamine (PPD). It was used to analyze a total of 1238 facial and masseteric nerve biopsies. The technique was validated by comparing manual and semi-automated quantification of 129 (10.4%) randomly selected biopsies. The software-based method demonstrated a sensitivity of 94% and a specificity of 87%. Semi-automatic axon counting was significantly faster (p < 0.001) than manual counting. It took 1 hour and 47 minutes for all 129 biopsies (averaging 50 sec per biopsy, 0.04 seconds per axon). The counting process is automatic and does not need to be supervised. Manual counting took 21 hours and 6 minutes in total (average 9 minutes and 49 seconds per biopsy, 0.52 seconds per axon). Our method showed a linear correlation to the manual counts (R = 0.944 Spearman rho). Attempts have been made by several research groups to automate axonal load quantification. These methods often require specific hard- and software and are therefore only accessible to a few specialized laboratories. Our semi-automated axon quantification is precise, reliable and time-sparing using publicly available software and should be useful for an effective axon quantification in various human peripheral nerves.
Highlights
We developed a time-efficient semi-automated axon quantification method using freeware in human cranial nerve sections stained with paraphenylenediamine (PPD)
It is known that axonal loads in peripheral nerve reconstruction, such as brachial plexus surgery, are similar to those seen in facial nerve surgery[20]
Medical treatment of any kind performed on peripheral nerves requires a high degree of knowledge about their macro and microanatomy
Summary
We developed a time-efficient semi-automated axon quantification method using freeware in human cranial nerve sections stained with paraphenylenediamine (PPD). Attempts have been made by several research groups to automate axonal load quantification These methods often require specific hard- and software and are only accessible to a few specialized laboratories. Marina et al have coined a method which, similar to this project, focuses on simpler semi-automated anaylsis[10] Other research groups such as Hunter et al focus on highly specialized methods, which are able to produce a wide range of data and process numerous variables[11]. The semi-automated quantification method proposed in this study was developed as part of a greater study on Human facial nerves, for which a time sparing, cost efficient and user-friendly method of axonal quantification was required. Quantifying axon capacity is a key part of the study, especially for determining a cut off axon count of 900 axons within a facial nerve branch
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