Diffusion tensor MRI (DT-MRI) is the only non-invasive method for characterising the microstructural organization of tissue in vivo. Generating parametric maps that help to visualise different aspects of the tissue microstructure (mean diffusivity, tissue anisotropy and dominant fibre orientation) involves a number of steps from deciding on the optimal acquisition parameters on the scanner, collecting the data, pre-processing the data and fitting the model to generating final parametric maps for entry into statistical data analysis. Here, we describe an entire protocol that we have used on over 400 subjects with great success in our laboratory. In the 'Notes' section, we justify our choice of the various parameters/choices along the way so that the reader may adapt/modify the protocol to their own time/hardware constraints.