Abstract: The study of Microfluidic chip has been a popular research area for its low reagent consumption, integration and high throughput. Researchers prefer glass chip as their experimental platform for its favourable optical properties and mature process. In this paper, the progress of wet-etchingin glass microfluidic chip was analyzed and the biocompatibilities of the substrate after etching technique were explored. The results show that, the broadening can reach 100 μm and biocompability is more reliable after H₂SO₄ disposed. As a result, it is necessary to provide appropriate cleaning and coating which might ensure follow-up operation. It provides a reference and guidance for microfluidic chip experiments.