Trypanosoma brucei lives an entirely extracellular life cycle in its mammalian host, facing a constant onslaught of host antibodies. The parasite evades clearance by the host immune system through antigenic variation of its dense variant surface glycoprotein (VSG) coat, periodically "switching" expression of the VSG using a large genomic repertoire of VSG-encoding genes. Studies of antigenic variation in vivo have focused exclusively on parasites in the bloodstream, but recent work has shown that many, if not most, parasites are extravascular and reside in the interstitial spaces of tissues. However, it is unknown whether parasites undergo antigenic variation while in extravascular spaces. We sought to explore the dynamics of antigenic variation in extravascular parasite populations using VSG-seq, a high-throughput sequencing approach for profiling VSGs expressed in populations of T. brucei. Our experiments show that the expressed VSG repertoire is not uniform across populations of parasites within the same infection and that a greater number of VSGs are expressed in tissue spaces than in the blood. More than 75% of the VSGs detected in an animal were found exclusively within extravascular spaces. Interestingly, we also noticed a delay in the VSG-specific clearance of parasites in tissue spaces compared to the blood. This finding aligns with a model in which parasites "hide" from the immune system in tissue spaces, where a slower immune response provides them with more time to generate new antigenic variants. Overall, our results show that extravascular spaces are significant reservoirs of VSG diversity, potentially resulting from delayed clearance of tissue-resident parasites.