To evaluate Henle's Fiber Layer (HFL) thickness and volume parameters in patients with cone photoreceptor atrophy with directional optical coherence tomography (D-OCT). Macular 20°×20° standard and D-OCT images were acquired from patients diagnosed with hereditary cone dystrophy with evident foveal ellipsoid zone defect in OCT, and age-matched healthy controls. Thickness and volume parameters of HFL, outer nuclear layer (ONL) and retinal layers between ellipsoid zone and Bruch membrane complex (EZ-BM) were calculated from manual segmentation through D-OCT images and comparative analysis is performed. Twelve eyes of 6 patients were compared to 12 eyes of 6 age-matched healthy controls (mean age: 29.5±16.6 and 26.6±3.9 years respectively; P=0.162). Patients had lower total HFL volume (0.45±0.03 and 0.85±0.15 mm3; P<0.001) and mean HFL thickness (16.1±1.1 and 30.1±5.3 µm; P<0.001) than healthy controls. Central subfield, parafoveal and perifoveal ETDRS zone HFL parameters in patients were significantly lower than healthy controls. A centrifugal correlation was found between central ONL and the corresponding parafoveal HFL (Spearman's rho: 0.785; P<0.001). HFL assessment might be a useful OCT biomarker in patients with cone photoreceptor atrophy. HFL thinning is observed in foveal, parafoveal and perifoveal areas of patients with cone photoreceptor atrophy, while volume reduction in ONL and EZ-BM components were limited to central and parafoveal zones.