A simple method using spin-deposition and sacrificial layer etching is used to fabricate all-polyimide nanochannels (100 and 500 nm channel height). Channels are characterized using spontaneous capillary filling with water, ethanol and isopropanol, and with electroosmotic flow. The channels can be produced with simple cleanroom equipment, namely spinning and metal deposition facilities. Polyimide is an excellent material for micro- and nanofluidic channels due to its favourable electrical and mechanical properties and its biocompatibility.