Fourier Transform methods were used to quantify mean elongation, mean orientation, and standard deviation of orientations of cultured vascular endothelial cells. Images of cell populations, which had been subjected to 11 and 20 hours of shear stress at 30 dynes/cm2 and 20 hours of no shear, were analyzed by Fourier Transform methods. Measurements of cell morphology and orientation characteristics were also obtained using a manual method for comparison purposes. The results of the study showed that mean cell orientation can be determined accurately with the Fourier Transform methods. Attempts to determine the standard deviation of cell orientations, however, resulted in poorer estimates of mean elongation and standard deviation of orientations except in the case of exposure of endothelial cells to 20 hours of shear, where the actual standard deviation of orientations was low. When the value for standard deviation of orientations was constrained to zero, a minimum possible mean elongation was determined reliably using the Fourier Transform methods. Use of the Fourier Transform methods in determining morphological and orientation characteristics of cell monolayers is fast and objective and may provide a basis for identifying other characteristics of cell shape.