In this chapter an overview of manufacturing methods, leading to the fabrication of microstructures in glass substrates, is presented. Glass is a material of excellent optical properties, a very good electric insulator, biocompatible and chemically stable. In addition to its intrinsic qualities, glass can be processed with the use of manufacturing methods originating from the microelectronic industry. In this text two complete manufacturing protocols are described, each composed of standard microfabrication steps; namely, the deposition of masking layers, photolithographic patterning and pattern transfer via wet or dry etching. As a result, a set of building blocks is provided, allowing the manufacture of various microfluidic components that are frequently used in the domain of micro-total analysis system technology.