ObjectivesTo compare choroidal vascularity index (CVI) measurements using the automated image binarization algorithm in healthy subjects with two Spectralis spectral-domain optical coherence tomography (SD-OCT) protocol scans.MethodsSixty-nine eyes of 69 healthy volunteers were included in this cross-sectional prospective study. Two subsequent horizontal 20°line scans passing through the fovea were acquired with enhanced-depth imaging mode with high speed (HS) and high resolution (HR) protocol scans. CVI and its subcomponents were measured with the previously validated automated algorithm. Differences between choroidal measurements obtained with HS and HR protocol scans were evaluated with t-test and Bland & Altman plots.ResultsA total of 33 male (47.8%) and 36 female (52.2%) subjects with a mean age of 35.1 ± 13.4 years were included. Overall, HS protocol scan was associated with significant lower values of total choroidal area (−0.047 mm2) and stromal choroidal area (−0.036 mm2), and a significant greater value of CVI (+0.010%) if compared to HR protocol. Luminal choroidal area was lower when calculated with the HS protocol, although it did not reach significance. To compare the two different protocols, the number of pixels should be multiplied for 3.87 ×5.73 when the CVI is measured on a HR OCT b scan and 3.87 ×11.46 for the HS OCT b scan.ConclusionsHS and HR acquisition modes significantly influence CVI and its subcomponents values measured with the automated software. However, adopting the scale factors can minimize the differences between the two protocol scans.