Background: Changes in choroidal thickness are associated with various ocular diseases and the choroid can be imaged using spectral-domain optical coherence tomography (SDOCT) and enhanced depth imaging OCT (EDIOCT). New Method: Eighty macular SDOCT volumes from 80 patients were obtained using the Zeiss Cirrus machine. Eleven additional control subjects had two Cirrus scans done in one visit along with EDIOCT using the Heidelberg Spectralis machine. To automatically segment choroidal layers from the OCT volumes, our graph-theoretic approach was utilized. The segmentation results were compared with reference standards from two graders, and the accuracy of automated segmentation was calculated using unsigned to signed border positioning thickness errors and Dice similarity coefficient (DSC). The repeatability and reproducibility of our choroidal thicknesses were determined by intraclass correlation coefficient (ICC), coefficient of variation (CV), and repeatability coefficient (RC). Results: The mean unsigned to signed border positioning errors for the choroidal inner and outer surfaces are 3.39plusminus1.26microns (mean plusminus SD) to minus1.52 plusminus 1.63microns and 16.09 plusminus 6.21microns to 4.73 plusminus 9.53microns, respectively. The mean unsigned to signed choroidal thickness errors are 16.54 plusminus 6.47microns to 6.25 plusminus 9.91microns, and the mean DSC is 0.949 plusminus 0.025. The ICC (95% CI), CV, RC values are 0.991 (0.977 to 0.997), 2.48%, 3.15microns for the repeatability and 0.991 (0.977 to 0.997), 2.49%, 0.53microns for the reproducibility studies, respectively. Comparison with Existing Method(s): The proposed method outperformed our previous method using choroidal vessel segmentation and inter-grader variability. Conclusions: This automated segmentation method can reliably measure choroidal thickness using different OCT platforms.