The purpose of this work was to empirically study the effect of two different functional MRI (fMRI) experimental design parameters on blood oxygenation level dependent (BOLD) signal contrast. The design parameters examined were: (1) the pattern of administering the control and activation conditions; and (2) the quantity of the control signals collected. Six normal volunteers underwent single finger opposition fMRI using T2*-weighted echo planar imaging technique. The BOLD signal contrast was evaluated using analysis of variance on normalized t-values and mean region-of-interest (ROI) intensity. The results demonstrate: (1) the alternating pattern of administering rest and activation epochs produced a more robust statistical difference in BOLD signal than a continuous pattern, and (2) a reduction in the amount of rest signal collected did not result in a statistically significant reduction in the BOLD signal. The results indicate that manipulating such factors of signal collection does alter BOLD signal properties.