Alternate　hot　and　cold　electrodes,　actively　heated　and　chilled,　were　facilely　fabricated　for　the　first　time.　It　consists　of　two　symmetrically　placed　semiconductor　thermoelectric　coolers　and　an　embedded　interlayer　with　a　Pt-PtRh　thermocouple.　The　tip　of　Pt-PtRh　thermocouple　can　be　used　as　both　an　electrochemical　electrode　and　a　temperature-sensing　device　simultaneously.　Theoretical　simulation　shows　that　a　local　steady-state　supercooled　water　within　a　radius　of　ca.　12.6　mm　around　the　electrode　can　be　obtained.　In　this　paper,　the　activity　of　horseradish　peroxidase　is　evidenced　at　supercooled　temperature　(-　9　°C)　in　0.1　M　PBS.　©　2015　Elsevier　B.V.　All　rights　reserved.