水冷式連續流聚合酶連鎖反應晶片系統之研製

傳統生化實驗中，檢測樣本必須恆溫培養於培養皿中，實驗完成後，過多的培養液或樣本必須拋棄。為節省成本和系統微小化，本研究目的是研發一個水冷式微域加熱晶片。以軟體CFD-ACE+TM模擬設計加熱晶片，晶片上設計兩個不同溫度的恆溫加熱區，中間為一冷卻水道，以水道裡的流體之流速改變，模擬晶片表面溫度的變化。模擬結果顯示，當流體在水道中的流速越快，水道的對流效應較強，晶片中央的溫度也會較低。在模擬中找出最佳流速控制，應用於實際晶片上。實驗方面則是以微機電製程和傳統加工方法製作晶片，水冷式微域加熱晶片可分成三個部份，包含PDMS和玻璃之晶片本體、加熱模組及冷卻水道。晶片具有供檢測樣本流動的PDMS流道，晶片下方兩側的加熱模組則是兩條可被熱控制的加熱鋁塊，且兩加熱器間夾有一個冷卻水道可使流體流動。當加熱器上方形成兩個不同溫度的恆溫區時，利用流體在冷卻水道中流動達到冷卻效果，使晶片表面呈現三個不同溫度的大面積恆溫區。實驗結果顯示，不同流體速度對晶片表面溫度影響很大，流量增大造成水冷式流道具有較大的熱對流效應，冷卻的效果很明顯。並且可在晶片上建立特定恆溫區，使其可以進行聚合酶鏈鎖反應實驗或細胞培養等實驗。

關鍵字

水冷式；加熱晶片；恆溫區；聚合酶連鎖反應

並列摘要

Traditionally, the biochemical samples are cultured in Petri dishes. After completing the experiment, excessively used sample must be disposed. In order to save the cost and miniaturize the system, this research is to develop a micro-domain heating chip with waterwall cooling channel. The commercial software CFD-ACE+TM is utilized to simulate the thermal fields of the chip. There are two different constant temperature regions designed within the chip. Between these two isothermal areas, there is a waterwall cooling channel and the fluid in the channel can be used to control the temperature of the chip. The effects of various fluid flow velocities on the temperature distribution are examined. Simulation result shows that when the fluid flow rate in the channel becomes slow, it will be more easily heated by the isothermal areas with high temperatures. This makes the fluid temperature near the outlet getting higher and then the surface temperature of the chip is non-uniform. The optimal flow rate is found and it can be apply to the experiment. In our experiment, the chip system mainly consists of three parts. They include a PDMS-glass chip, two heating module and the fluid channel used to control the chip temperature. A PMMA channel for fluid flowing is fabricated under the chip. The heating module comprises two heaters that can be thermal controlled. When two different isothermal regions are created at the chip surface, we make use of the fluid which flows in the waterwall channel between the two heaters in order to cool the chip. Then three different isothermal zones can be created on the chip. The result shows that the fluid is easily heated by isothermal area, the effects of various flow rate in the channel on the surface temperature is noticeable. The obvious heat convective effect inside the cooling channel can be observed with the increasing of the fluid velocities. The large areas of the various isothermal regions are shown at the chip surface. Experiment results are compared with the simulated data, and it shows a similar trend. In the future we may establish several isothermal areas in the chip, and it could be applied in the fields of cell culture, drug screening or polymerase chain reaction in the chips.

並列關鍵字

Waterwall cooling ； Heating chip ； Isothermal region ； Polymerase chain reaction

參考文獻

[1] Yao, L., Liu, B., Chen, T., Liu, S., and Zuo, T., 2005, “Micro flow-through PCR in a PMMA chip fabricated by KrF excimer laser,” Biomedical Microdevices, Vol. 7, No. 3, pp. 253-257.

[2] Xiang, Q., Xu, B., Fu, R., and Li, D., 2005, “Real time PCR on disposable PDMS chip with a miniaturized thermal cycler,” Biomedical Microdevices, Vol. 7, No. 4, pp. 273-279.

[3] Consolandi, C., Severgnini, M., Frosini, A., Caramenti, G., Fazio, M. D., Ferrara, F., Zocco, A., Fischetti, A., Palmieri, M., and Bellis, G. D., 2006, “Polymerase chain reaction of 2-kb cyanobacterial gene and human anti-α1-chymotrypsin gene from genomic DNA on the In-Check single-use microfabricated silicon chip,” Analytical Biochemistry, Vol. 353, No. 2, pp. 191-197.

[4] Xiang, Q., Xu, B., and Li, D., 2007, “Miniature real time PCR on chip with multi-channel fiber optical fluorescence detection module,” Biomedical Microdevices, Vol. 9, No. 4, pp. 443-449.

[5] Mondal, S., and Venkataraman, V., 2007, “Novel fluorescence detection technique for non-contact temperature sensing in microchip PCR,” Journal of Biochemical and Biophysical Methods, Vol. 70, No. 5, pp. 773-777.

國際替代計量

水冷式連續流聚合酶連鎖反應晶片系統之研製

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