Objective: To understand microstructural changes after myocardial infarction (MI), we evaluated myocardial fibers of rhesus monkeys during acute or chronic MI, and identified the differences of myocardial fibers between acute and chronic MI. Materials and Methods: Six fixed hearts of rhesus monkeys with left anterior descending coronary artery ligation for 1 hour or 84 days were scanned by diffusion tensor magnetic resonance imaging (MRI) to measure apparent diffusion coefficient (ADC), fractional anisotropy (FA) and helix angle (HA). Results: Comparing with acute MI monkeys (FA: 0.59 +/- 0.02; ADC: 5.0 +/- 0.6 x 10(-4) mm(2)/s; HA: 94.5 +/- 4.4 degrees), chronic MI monkeys showed remarkably decreased FA value (0.26 +/- 0.03), increased ADC value (7.8 +/- 0.8 x 10(-4)mm(2)/s), decreased HA transmural range (49.5 +/- 4.6 degrees) and serious defects on endocardium in infarcted regions. The HA in infarcted regions shifted to more components of negative left-handed helix in chronic MI monkeys (-38.3 +/- 5.0 degrees-11.2 +/- 4.3 degrees) than in acute MI monkeys (-41.4 +/- 5.1 degrees-53.1 +/- 3.7 degrees), but the HA in remote regions shifted to more components of positive right-handed helix in chronic MI monkeys (-43.8 +/- 2.7 degrees-66.5 +/- 4.9 degrees) than in acute MI monkeys (-59.5 +/- 3.4 degrees-64.9 +/- 4.3 degrees). Conclusion: Diffusion tensor MRI method helps to quantify differences of mechanical microstructure and water diffusion of myocardial fibers between acute and chronic MI monkey's models.