Background: global longitudinal strain (GLS) measures myocardial deformation and is a sensitive modality for detecting subclinical myocardial dysfunction and predicting cardiac outcomes. The accuracy of speckle-tracking echocardiography (STE) is dependent on temporal resolution. A novel software enables relatively high frame rate (Hi-FR) (~200 fps) echocardiographic images acquisition which empowers us to investigate the impact of Hi-FR imaging on GLS analysis. The goal of this pilot study was to demonstrate the feasibility of Hi-FR for STE. Methods: In this prospective study, we acquired echocardiographic images using clinical scanners on patients with normal left ventricular systolic function using Hi-FR and conventional frame rate (Reg-FR) (~50 FPS). GLS values were evaluated on apical 4-, 2- and 3-chamber images acquired in both Hi-FR and Reg-FR. Inter-observer and intra-observer variabilities were assessed in Hi-FR and Reg-FR. Results: There were 143 resting echocardiograms with normal LVEF included in this study. The frame rate of Hi-FR was 190 ± 25 and Reg-FR was 50 ± 3, and the heart rate was 71 ± 13. Strain values measured in Hi-FR were significantly higher than those measured in Reg-FR (all p < 0.001). Inter-observer and intra-observer correlations were strong in both Hi-FR and Reg-FR. Conclusions: We demonstrated that strain values were significantly higher using Hi-FR when compared with Reg-FR in patients with normal LVEF. It is plausible that higher temporal resolution enabled the measurement of myocardial strain at desired time point. The result of this study may inform clinical adoption of the novel technology. Further investigations are necessary to evaluate the value of Hi-FR to assess myocardial strain in stress echocardiography in the setting of tachycardia.