The recent drive in biomedical research has been mainly focused on detection, diagnosis, treatment, and prevention of diseases in order to ultimately foster better health. Detection of cancerous tissue or relevant cancer- causing entities using an extremely low sample volume and at an early stage is another targeted objective of researchers in the recent past. From that perspective, ultrasensitive detection and imaging methods are treated as enablers for the advancement of diagnostic methodologies to the next level. The steady progress in biology, engineering, and medicine toward diagnostics based on molecular markers leads to the exploration of high-throughput methods for the detection of biomolecules and their interactions in a biological system. Fluorescence-based bioassays are novel diagnostic tools available to clinicians for deciding on further treatment and to researchers for monitoring biological functions that lead to novel investigations.