Genome sequencing methods have led to a rapid expansion in our understanding of the subtleties and variations in how genetic profiles affect disease. This has been spurred in part by new technological advances. Nanopore sequencing has been highlighted as one upcoming technology with the potential to go further, and improve the speed and cost effectiveness of seqencing. In nanopore sensing, the flow of ions through a nanoscopic protein or solid-state pore is disrupted by the presence of an analyte. If this analyte is DNA, blockades in the current corresponding to the sequence are detected. Long reads of many thousands of bases can be made without amplification or labelling. Our research track record marks a long-standing interest in developing methods capable of detecting individual molecules using fluorescence microscopy. We have used this to improve our understanding of membrane protein function, and in particular, how specialized protein pores work. Most recently we used these methods to demonstrate DNA base detection in a nanopore using an optical, rather than an electrical readout. These methods have the potential to improve the parallelization of nanopore sensing. In collaboration with Oxford Nanopore Technologies we have identified two essential challenges for current nanopore sensing where an optical readout provides significant impact: 1. We will combine optical single channel recording and single-molecule fluorescence imaging to understand and then optimise the mechanistic steps of nanopore sensing; 2. We will go beyond the current speed and sensitivity limitations of optical single channel recording and develop new single-molecule microscopy methods for nanopore sensing.