The Hsp70 family of molecular chaperones represents a central hub of the proteostasis network, performing a plethora of cellular roles including de novo protein folding and refolding. The mechanisms by which Hsp70 folds client proteins are highly regulated by its co-chaperones Hsp40 and nucleotide-exchange factors (NEFs). Whilst the ability of Hsp70 molecular chaperones to remodel the conformation of their clients is central to their biological function, questions remain regarding the precise molecular mechanisms by which the Hsp70 chaperone machinery interacts with the client and how this contributes towards efficient protein folding. Here, we describe our work using Total Internal Reflection Fluorescence (TIRF) microscopy and single-molecule Fluorescence Resonance Energy Transfer (smFRET) to temporally observe the conformational changes that occur to individual firefly luciferase (Fluc) proteins as they are folded by the Hsp70 system. For the first time, we observed multiple cycles of chaperone binding-and-release to an individual client during refolding and that high rates of chaperone cycling improves refolding yield. Furthermore, we demonstrate that Hsp40 remodels misfolded proteins via a conformational selection mechanism whereas Hsp70 resolves misfolded states via mechanical unfolding. This work illustrates that the temporal observation of chaperone-assisted folding enables the elucidation of key mechanistic details inaccessible using other approaches.