provides information about molecular vibrations, which can be used for molecular detection, identification, and diagnosis. [1] Over the last few decades, surface-enhanced Raman scattering (SERS) techniques, in which the Raman signals are significantly enhanced based on strong electromagnetic fields amplified by noble metal nanostructures, have been developed, demonstrating powerful analytic performance, including superb sensitivity for the detection of molecules. [2][3][4] SERS-based detection has been explored in a broad range of research fields as an attractive and promising spectroscopic technique. In particular, microfluidic devices coupled with SERS have been recently demonstrated for in situ monitoring of chemical reactions and detection of analytes. [5][6][7] To obtain SERS-integrated microfluidic devices, SERS-active metallic nanostructures fabricated by various methods, such as electromigration, [8] E-beam lithography, [9,10] A novel droplet-based surface-enhanced Raman scattering (SERS) sensor for highthroughput real-time SERS monitoring is presented. The developed sensors are based on a droplet-guiding-track-engraved superhydrophobic substrate covered with hierarchical SERS-active Ag dendrites. The droplet-guiding track with a droplet stopper is designed to manipulate the movement of a droplet on the superhydrophobic substrate. The superhydrophobic Ag dendritic substrates are fabricated through a galvanic displacement reaction and subsequent self-assembled monolayer coating. The optimal galvanic reaction time to fabricate a SERS-active Ag dendritic substrate for effective SERS detection is determined, with the optimized substrate exhibiting an enhancement factor of 6.3 × 10 5 . The height of the droplet stopper is optimized to control droplet motion, including moving and stopping. Based on the manipulation of individual droplets, the optimized droplet-based real-time SERS sensor shows high resistance to surface contaminants, and droplets containing rhodamine 6G, Nile blue A, and malachite green are successively controlled and detected without spectral interference. This noble droplet-based SERS sensor reduces sample preparation time to a few seconds and increased detection rate to 0.5 µL s −1 through the simple operation mechanism of the sensor. Accordingly, our sensor enables high-throughput real-time molecular detection of various target analytes for real-time chemical and biological monitoring.