Abstract: Static chip polymerase chain reaction(PCR) needs no mechanical parts, and is easy to be integrated with chip separation and detection system, thus has a wider application in analysis field and POCT(point of care test). Chip thermal-time program control is crucial in static chip PCR, normally this is achieved with the aid of micro-fabrication technique in preparing the heating and sensing elements on chip. In this article, a thermostat chip that is capable of self-heating and sensing automatically and programmatically was established for the first time with commercial indium-tin oxide(ITO) glass substrate without involving any micro-fabrication process. The transparent and elelctro-conductive layer was used both as heating and temperature sensing elements, and time delay due to separate sensor is minimized. Electrical contacts is reduced from two couples(one for heating and one for sensing) to only one for each thermostat chip unit. The system has a control stability of 0.2 ℃, a maximum temperature ramping rate above 20 ℃/s and a cooling rate 8 ℃/s with the aid of a cooling fan. Multiple open reaction reservoirs can be constructed directly on the chip surface, and static chip PCR of a λDNA fragment(c.a. 157 bp) is successfully demonstrated. In-situ real-time fluorescence monitoring of a dsDNA and SYBR Green Ⅰ mixture during the temperature cycling was also demonstrated by placing the chip on the sample stage of an inverted fluorescence microscope that equipped with a blue light emission diode(575 nm) and a photomultiplier tube.

Key words: Indium tin oxide(ITO) glass, Self-heating and sensing temperature control, Static chip PCR, In-situ real time fluorescence monitoring, Light emission diode