Functionally targeted assemblies are of rapidly emerging importance for diagnostic and therapeutic applications since they maximize delivered dosage of contrast agents and drugs to select targets. This is an experimental Biophysics study which will develop an optimal method for the detection of DNA using near-infrared (NIR) emissive polymersomes. NIR emissive polymersomes with the specific adhesive properties have the enormous potential for non-invasive in vivo biomedical imaging and diagnostics of diseases. In this study, however, we use polymersomes linked with single-stranded DNA to bind to DNA coated surfaces. This specific adhesion of NIR emissive polymersomes to substrates is driven by DNA hybridization induced by large numbers of specific hybridization associating single-stranded DNA molecules bound to their surface. The targeted adhesion of these NIR emissive polymersomes is probed and measured by optical videomicroscopy (micron-sized) and through their multiplexed emissive signals from NIR emitting porphyrin dyes incorporated into the polymersomes (micron and nano-sized). This simple model allow the development of the essential design principles for targeting NIR emissive polymersomes to any substrates, at the same time, enable real-time detection and identification of limited targets.