Magnetic Resonance Imaging (MRI) has come to be known as a unique radiological imaging modality because of its ability to perform tomographic imaging of body without the use of any harmful ionizing radiation. The radiologists use MRI to gain insight into the anatomy of organs, including the brain, while biomedical researchers explore the modality to gain better understanding of the brain structure and function. However, due to limited resolution and contrast, the conventional MRI fails to show the brain microstructure. Diffusion Tensor Imaging (DTI) harnesses the power of conventional MRI to deduce the diffusion dynamics of water molecules within the tissue and indirectly create a three-dimensional sketch of the brain anatomy. DTI enables visualization of brain tissue microstructure, which is extremely helpful in understanding various neuropathologies and neurodegenerative disorders. In this review, we briefly discuss the background and operating principles of DTI, followed by current trends in DTI applications for biomedical and clinical investigation of various brain diseases and disorders.