BACKGROUND: Cardiovascular magnetic resonance based tissue tracking (CMR-TT) was reported to provide detailed insight into left ventricular (LV) contractile function and deformation with both of two- and three-dimensional (2/3D) algorithms. This study was designed to investigate the feasibility and reproducibility of these two techniques for measuring LV global and segmental strain, and establish gender- and age-related reference values of global multi-dimensional peak strains among large healthy population. METHODS: We retrospectively recruited 150 healthy volunteers (75 males/females) and divided them into three age groups (G(20-40), G(41-60) and G(61-80)). LV global mean and peak strains as well as segmental strains in radial, circumferential and longitudinal directions were derived from post-hoc 2/3D CMR-TT analysis of standard steady-state free precession (SSFP) cine images acquired at 1.5T field strength. RESULTS: Both 2D and 3D CMR-TT modalities enable the tracking of LV myocardial tissues and generate global and segmental strain data. By comparison, 3D CMR-TT was more feasible in measuring segmental deformation since it could generate values at all segments. The amplitudes of LV 3D global peak strain were the smallest among those of 2/3D corresponding global mean or peak strains except in the radial direction, and was highly correlated with 2D global mean strains (correlation coefficient r=0.71–0.90), 2D global peak strains (r=0.75–0.89) and 3D global mean strains (all r=0.99). In healthy cohort, LV 3D global peak values were 44.4%±13.0% for radial, −17.0%±2.7% for circumferential and −15.4%±2.3% for longitudinal strain. Females showed significantly larger amplitude of strains than males, especially in G(61-80) (P