64Cu is a cyclotron-produced radionuclide which offers, due to its characteristic decay scheme the possibility to combine PET investigations with radiotherapy. We evaluated the Alceo sys-tem from Comecer SpA to automatically produce 64Cu for radiolabelling purposes. We established a 64Cu production routine with high yields and radionuclide purity in combination with excellent operator radiation protection. The carbonic anhydrase XII targeting 6A10 antibody fab fragment was successfully radiolabelled with the produced 64Cu and proof of principle small animal PET experiments on mice bearing glioma xenografts were performed. We obtained a high tumor-to-contralateral muscle ratio, which encourages further in vivo investigations of the radioconjugate regarding a possible application in diagnostic tumor imaging.