Our aim was to provide, for the first time, reference thickness values for the SD-OCT posterior pole algorithm (PPA) available for Spectralis OCT device (Heidelberg Engineering, Heidelberg, Germany) and to analyze the correlations with age, gender and axial length. We recruited 300 eyes of 300 healthy Caucasian subjects between 18 and 84 years. By PPA, composed of 64 (8 × 8) cells, we analyzed the thickness of the following macular layers: retinal nerve fiber layer (RNFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), retinal pigment epithelium (RPE), inner retina, outer retina and full retina. Mean ± SD, 1st, 5th, 95th percentiles were obtained for each cell at all macular layers. Significant negative correlations were found between age and thickness for most macular layers. The mean thickness of most macular layers was thicker for men than women, except for RNFL, OPL and RPE, with no gender differences. GCL, IPL and INL thicknesses positively correlated with axial length in central cells, and negatively in the cells near the optic disk. The mean RNFL thickness was positively associated with axial length. This is the first normative database for PPA. Age, gender and axial length should be taken into account when interpreting PPA results.