Glass is one of the most ideal materials for micro/nanofluidic devices due to its excellent optical transparency, resistance to a wide range of solvents and reagents, and easy to modify surfaces by silane-coupling reagents. From a practical point of view, glass is a hard material and is suitable for real applications. One of the advantages of glass is its reusability; however, this reusability is difficult to realize in certain conditions. Washing or re-modification of micro/nanofluidic channels is sometimes difficult due to the ultrasmall space in these channels. If the glass devices are detachable, it is easy to access the channel surface, and the channels can be cleaned and re-modified. When the substrates are bonded again, the devices are fabricated easily without repeating laborious and expensive micro/nano-fabrication processes. This technology gives researchers and users a choice of glass substrates in fundamental research studies and real-time applications. In this study, we propose a detachable glass micro/nanofluidic device by our low temperature bonding method. The surface bonding energy is controlled to realize both high pressure capacity for micro/nanofluidics and easy separation of glass substrates without fracturing. As a result, at least four times detaching and bonding is confirmed.