Purpose: To assess the reproducibility of peripapillary retinal nerve fiber layer (RNFL) thickness, measured with Spectralis® OCT, in healthy and glaucomatous subjects, through of inter-examiner variability, intra-test and inter-test with and without software of followup. Methods: Cross-sectional and prospective study with 37 controls (63 eyes) and 33 patients with glaucomatous optic neuropathy (55 eyes). Intraclass correlation coefficient (ICC) and coefficient of variation (CV) were calculated. Results: Low inter-examiner, intra-test and inter-test variability, with ICC>0,75 e CV