Detection of multiple relaxation pools using MRI is useful in a number of neuro-pathologies including multiple sclerosis (MS), Alzheimer's, and stroke. In this study we evaluate the feasibility of using T1ρ imaging for the detection of bi-exponential decays in the human brain. A prospective T1ρ imaging study was performed on model relaxation phantoms (eggs) and 7 healthy volunteers. The data was fitted using a single pool and a 2-pool model to estimate mono- and bi-exponential T1ρ maps, respectively. Bi-exponential decays were identified in the gray matter (GM) and white matter (WM) of the brain with 40.5% of GM, and 65.1% of WM pixels showing two T1ρ relaxation pools (significance level P < 0.05). Detection of T1ρ based bi-exponential decays in the brain provides complimentary information to T2 based contrast regarding the in vivo micro-environment in the brain.