One of the most significant challenges facing the healthcare community is the accurate and selective detection and quantification of trace levels of analyte. This can have enormous impact with numerous application, especially in the context of biosensing, where applications can range from early stage detection of disease to the identification of new markers amongst others. These challenges are compounded when the analysis of real samples or even complex mixtures are required. To address this need, single molecule or near single molecule methods are currently being heavily pursued which allows for the discrimination of events with low copy numbers by removing the “clouding” associated with ensemble averaging. Although strategies for detecting single molecules have existed for a number of years, efficient and cost-effective label-free methods without the need for chemical modification are lacking especially when considering smaller molecules directly in unprocessed clinical samples. As part of this proposal, I intend to address this limitation by developing new strategies in single molecule nanoscale sensing which will be both highly sensitive and selective for improved therapeutics and diagnostics applications.