Sweat loss monitoring is important for understanding the body’s thermoregulation and hydration status, as well as for comprehensive sweat analysis. Despite recent advances, developing a low-cost, scalable, and universal method for the fabrication of colorimetric microfluidics designed for sweat loss monitoring remains challenging. In this study, we propose a novel laser-engraved surface roughening strategy for various flexible substrates. This process permits the construction of microchannels that show distinct structural reflectance changes before and after sweat filling. By leveraging these unique optical properties, we have developed a fully laser-engraved microfluidic device for the quantification of naked-eye sweat loss. This sweat loss sensor is capable of a volume resolution of 0.5 μL and a total volume capacity of 11 μL, and can be customized to meet different performance requirements. Moreover, we report the development of a crosstalk-free dual-mode sweat microfluidic system that integrates an Ag/AgCl chloride sensor and a matching wireless measurement flexible printed circuit board. This integrated system enables the real-time monitoring of colorimetric sweat loss signals and potential ion concentration signals without crosstalk. Finally, we demonstrate the potential practical use of this microfluidic sweat loss sensor and its integrated system for sports medicine via on-body studies.