We introduce a method to determine the retinal nerve fiber layer (RNFL) thickness in OCT images based on anisotropic noise suppression and deformable splines. Spectral-Domain Optical Coherence Tomography (SDOCT) data was acquired at 29 kHz A-line rate with a depth resolution of 2.6 m and a depth range of 1.6 mm. Areas of 9.6x6.4 mm(2) and 6.4x6.4 mm(2) were acquired in approximately 6 seconds. The deformable spline algorithm determined the vitreous-RNFL and RNFL-ganglion cell/inner plexiform layer boundary, respectively, based on changes in the reflectivity, resulting in a quantitative estimation of the RNFL thickness. The thickness map was combined with an integrated reflectance map of the retina and a typical OCT movie to facilitate clinical interpretation of the OCT data. Large area maps of RNFL thickness will permit better longitudinal evaluation of RNFL thinning in glaucoma. (c) 2005 Optical Society of America.