This paper presents a new research study on the platform fabrication of fluorescence bio-detection chip with an optical fiber transmission. Anisotropic wet etching on (100) silicon wafers to fabrication V-groove for optical fiber alignment and micro-mirror were included. Combing with anodic bonding technique to adhere glass, silicon structure and optical fiber for a fluorescence excitation platform was completed. In this study, the etching solution 40% KOH was used to study the parameters effect. The results show that working temperature is the main parameter to significantly effect the etch rate. The anisotropic etching resulted 54.7° reflective mirrors and its reflectivity for optical beam were also examined. The surface roughness of the micromirror is Ra 4.1 nm measured using AFM, it provides excellent optical reflection. The incident light and beam profiles were also examined for further study. This study can show this micro-platform adaptable for fluorescence bio-detection.