ASSESSMENT AND IMPROVEMENT OF THE THERMAL PERFORMANCE OF A POLYCARBONATE MICRO CONTINUOUS FLOW POLYMERASE CHAIN REACTOR (CFPCR)

摘要

BioMEMS are compact devices that use microfabrication to miniaturize benchtop instrumentation. Due to the requirement for uniform temperature distributions over restricted areas, thermal isolation, and faster heating and cooling rates in a limited space, thermal management is a key to ensuring successful performance of BioMEMS devices. The continuous flow polymerase chain reactor (CFPCR) is a compact BioMEMS device that is used to amplify target DNA fragments using repeated thermal cycling. The temperature distribution on the backside of a micro CFPCR was measured using thermochromic liquid crystals and an infrared camera. In the liquid crystal experiment, the performance of a 5 mm thick polycarbonate micro CFPCR with thin film heaters attached directly to the bottom polycarbonate surface over each temperature zone was studied. Natural convection was used as a cooling mechanism. The temperature distribution in the renaturation zone was dependent on the positions of the feedback thermocouples in each zone. Three different thermocouple configurations were assessed and the liquid crystal images showed that a best case 3.86°C temperature difference across the zone, leading to a 20% amplification efficiency compared to a commercial thermal cycler [5]. The device was modified to improve the temperature distribution: a thinner substrate, 2 mm, reduced the thermal capacitance; grooves were micro-milled in the backside to isolate each temperature zone; and three separate copper heating stages, combining the thin film heaters with copper plates, applied uniform temperatures to each zone [10]. Infrared camera images showed that the temperature distributions were distinct and uniform with a ±0.3°C variations in each temperature zone, improving amplification efficiency to 72%. Good thermal management for PCR amplification can't only increase its reliability and yield efficiency, but also accelerate the entire analytical process.