Heat shock proteins (Hsps) are a family of ubiquitous molecular chaperones that are up-regulated under stress conditions. Hsps have a diverse range of cellular activities and play a vital role in proteostasis. A considerable amount of information concerning Hsps has been amassed over the past decades, yet many fundamental questions regarding the molecular mechanism(s) of Hsps remain unanswered. These questions are challenging to answer as these chaperones are notoriously difficult to study using traditional bulk analysis techniques, due to their dynamic nature. Over the past two decades, the use of single-molecule techniques to study biological systems has become more commonplace. By observing proteins one-by-one, rare and transient species can be characterised in great detail. Therefore the aims of this project are to use single molecule fluorescence microscopy to i) investigate the kinetics and stoichiometries of the interactions between Hsps and client proteins, and ii) identify how different members of the Hsps cooperate to prevent protein aggregation. We have characterised the ability of Hsps to prevent the aggregation of a novel model client using bulk assays. Furthermore, we have demonstrated that this interaction is observable at the single molecule scale. Overall, addressing these crucial gaps in knowledge will contribute to our understanding of the molecular mechanisms by which Hsps function as effective molecular chaperones.