Droplet-based transport driven by surface tension has been explored as an automated pumping source for several biomedical applications. This paper presented a simple and fast superhydrophobic modify and patterning approach to fabricate various open-surface platforms to manipulate droplets to achieve transport, mixing, concentration, and rebounding control. Several commercial reagents were tested in our approach, and the Glaco reagent was selected to create a superhydrophobic layer; laser cutters are utilized to scan on these superhydrophobic surface to create gradient hydrophilic micro-patterns. Implementing back-and-forth vibrations on the predetermined parallel patterns, droplets can be transported and mixed successfully. Colorimetry of horseradish peroxidase (HRP) mixing with substrates also reduced the reaction time by more than 5-times with the help of superhydrophobic patterned chips. Besides, patterned superhydrophobic chips can significantly improve the sensitivity of colorimetric glucose-sensing by more than 10 times. Moreover, all bioassays were distributed homogeneously within the region of hydrophilic micropatterns without the coffee-ring effect. In addition, to discuss further applications of the surface wettability, the way of controlling the droplet impacting and rebounding phenomenon was also demonstrated. This work reports a rapid approach to modify and patterning superhydrophobic films to perform droplet-based manipulations with a lower technical barrier, higher efficiency, and easier operation. It holds the potential to broaden the applications of open microfluidics in the future.