As in vivo flow behavior can be pulsatile, intermittent, and/or otherwise changeable with time, the ability to provide clinicians with a means of real-time visualization and functional assessment of structures is of particular importance. The discernment of pulsatile flow behavior using a dual-beam spectral domain optical coherence tomography system (db-SdOCT) by quasi-simultaneous measurement by two planes of illumination is demonstrated. By cross-correlation analysis, it is possible to compute velocity metrics pertaining to flowing particle motion, without a priori angular knowledge. This is the first application of cross-correlation-based dynamic assessment for the extraction of pulsatile behavior in an in vitro environment using an optimized db-SdOCT system. The experimental results outlined have shown the db-SdOCT system and its associated algorithms to be successful in the discernment of intermittent pulsatile flow behavior in in vitro models, concurrent to yielding velocity values in good agreement with that of the applied flow rate. (C) 2013 Society of Photo-Optical Instrumentation Engineers (SPIE)