Fluorometric sensors and devices are important tools for the detection and measurement of biomarkers, which are indicators of biological processes and diseases. These sensors involve the emission of light from fluorescent molecules upon interaction with target biomolecules, resulting in a measurable signal. Fluorescence-based techniques, for instance, fluorescence microscopy, fluorometry, fluorescence resonance energy transfer (FRET), and fluorescent probes, enable researchers to study biological molecules with high sensitivity and specificity. Fluorometric sensors offer numerous benefits over conventional detection methods, like higher sensitivity, selectivity, versatility, and the ability to sense various biomarkers instantaneously. In this chapter, we provide an overview of the recent developments in fluorometric sensors and devices for biomarker detection, including the design principles, sensing mechanisms, and fabrication techniques for developing fluorometric sensors. The challenges and future directions of this technology are also discussed. This chapter highlights the importance of these techniques in advancing our understanding of biological systems and their potential for improving healthcare, food safety, and environmental monitoring.