\　Single-cell　imaging,　a　powerful　analytical　method　to　study　single-cell　behavior,　such　as　gene　expression　and　protein　profiling,　provides　an　essential　basis　for　modern　medical　diagnosis.　The　coding　and　localization　function　of　microfluidic　chips　has　been　developed　and　applied　in　living　single-cell　imaging　in　recent　years.　Simultaneously,　chip-based　living　single-cell　imaging　is　also　limited　by　complicated　trapping　steps,　low　cell　utilization,　and　difficult　high-resolution　imaging.　To　solve　these　problems,　an　ultra-thin　temperature-controllable　microwell　array　chip　(UTCMA　chip)　was　designed　to　develop　a　living　single-cell　workstation　in　this　study　for　continuous　on-chip　culture　and　real-time　high-resolution　imaging　of　living　single　cells.　The　chip-based　on　ultra-thin　ITO　glass　is　highly　matched　with　an　inverted　microscope　(or　confocal　microscope)　with　a　high　magnification　objective　(100　xoil　lens),　and　the　temperature　of　the　chip　can　be　controlled　by　combining　it　with　a　home-made　temperature　control　device.　High-throughput　single-cell　patterning　is　realized　in　one　step　when　the　microwell　array　on　the　chip　uses　hydrophilic　glass　as　the　substrate　and　hydrophobic　SU-8　photoresist　as　the　wall.　The　cell　utilization　rate,　single-cell　capture　rate,　and　microwell　occupancy　rate　are　all　close　to　100%　in　the　microwell　array.　This　method　will　be　useful　in　rare　single-cell　research,　extending　its　application　in　the　biological　and　medical-related　fields,　such　as　early　diagnosis　of　disease,　personalized　therapy,　and　research-based　on　single-cell　analysis.　(C)　2021　Published　by　Elsevier　B.V.　on　behalf　of　Chinese　Chemical　Society　and　Institute　of　Materia　Medica,　Chinese　Academy　of　Medical　Sciences.