HIV incorporates its own protease which upon activation will transform the virus from immature to the infectious mature state. HIV also recruits cellular proteins termed Endosomal Sorting Complexes Required for Transport (ESCRTs) to facilitate its release from the host cell. We have shown that disrupting the balance between budding and protease activation will result in release of non infectious virus. Here we show how we use a variety of single molecule imaging as well as biochemical and virological tools to understand the molecular choreography between ESCRTs and HIV proteins during release of infectious virus.