The objectives of this study were (1) to investigate the passive stiffness of the medial gastrocnemius (MG) and lateral gastrocnemius (LG) in patients with and without plantar fasciitis (PF), (2) to explore the correlation between gastrocnemius stiffness and plantar fascia thickness (PFT) as well as the intensity of pain in patients with PF, (3) to detect optimal cut-off points for stiffness of the MG and LG for identifying patients with PF. Forty patients (mean age = 51.1 years ± 12.9) participated in this study. The elastic properties of the MG and LG were quantified using shear wave elastography ultrasound. The thickness of the plantar fascia was measured by B-mode imaging. The intensity of pain was assessed using a visual analogue scale. The results showed that when the ankle was in the relaxed position, patients with PF had increased passive stiffness in the MG (P < 0.05) but not in the LG. Significant correlations were found between pain and the stiffness of the MG (middle, distal; all P-values < 0.05) and no correlation was observed between pain and PFT (P = 0.416). The initial cut-off point for the stiffness of the MG was 29.08 kPa when the ankle was in the relaxed position. The findings from the present study show that an increase in muscle stiffness is not the same in the individual muscles of the gastrocnemius muscle. Traditional treatment of the whole gastrocnemius muscle might not be targeted at the tight muscle.