Wu P.-YWu J.-HYI-TING HSIEHChen L.C.-CCheng TWu P.-YHsieh B.-JHuang W.-LSHENG-LUNG HUANGWEI-LI CHEN2022-04-252022-04-25202120452322https://www.scopus.com/inward/record.uri?eid=2-s2.0-85115275779&doi=10.1038%2fs41598-021-97567-y&partnerID=40&md5=2fc7e8df707a00a4c1635bebb27e3f02https://scholars.lib.ntu.edu.tw/handle/123456789/607283This study aimed to evaluate the reliability of in vivo confocal microscopic neuroanalysis by beginners using manual and automated modules. Images of sub-basal corneal nerve plexus (SCNP) from 108 images of 18 healthy participants were analyzed by 7 beginner observers using manual (CCMetrics, [CCM]) and automated (ACCMetrics, [ACCM]) module. SCNP parameters analyzed included corneal nerve fiber density (NFD), corneal nerve branch density (NBD), corneal nerve fiber length (NFL), and tortuosity coefficient (TC). The intra-observer repeatability, inter-observer reliability, inter-module agreement, and left–right eye symmetry level of SCNP parameters were examined. All observers showed good intra-observer repeatability using CCM (intraclass correlation coefficient [ICC] > 0.60 for all), except when measuring TC. Two observers demonstrated especially excellent repeatability in analyzing NFD, NBD, and NFL using manual mode, indicating the quality of interpretation may still be observer-dependent. Among all SCNP parameters, NFL had the best inter-observer reliability (Spearman’s rank-sum correlation coefficient [SpCC] and ICC > 0.85 for the 3 original observers) and left–right symmetry level (SpCC and ICC > 0.60). In the additional analysis of inter-observer reliability using results by all 7 observers, only NFL showed good inter-observer reliability (ICC = 0.79). Compared with CCM measurements, values of ACCM measurements were significantly lower, implying a poor inter-module agreement. Our result suggested that performance of quantitative corneal neuroanalysis by beginners maybe acceptable, with NFL being the most reliable parameter, and automated method cannot fully replace manual work. ? 2021, The Author(s).adultarticlecontrolled studycorrelation coefficienteyefemalehumanhuman experimentin vivo studyinterrater reliabilitymajor clinical studymalemanual labornerve fibernerve plexusquantitative analysis[SDGs]SDG3journal article10.1038/s41598-021-97567-y345218902-s2.0-85115275779