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研究生:謝澤余
研究生(外文):Tse-Yu Hsieh
論文名稱:往復式熱循環反應器之研製
論文名稱(外文):Development of the Oscillatory Thermal Cycler Chamber
指導教授:陳志堅陳志堅引用關係
指導教授(外文):Jyh-Jian Chen
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:生物機電工程系所
學門:工程學門
學類:機械工程學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:90
中文關鍵詞:單晶片熱循環PCR接觸熱阻
外文關鍵詞:PCRPWMthermal cyclercontact thermal resistance
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論文摘要內容:本論文結合伺服馬達、8051 單晶片PWM 溫控裝置和以铣床加工之加熱機構整合成一個往復式熱循環裝置。以PCR 所需之溫度和反應所需之時間設計往復式熱循環裝置的加熱溫區,再以CFDRC 模擬軟體配合副程式撰寫的邊界條件,做最佳化設計。探討反應槽於熱循環溫區內移動速率對反應槽溫度的影響,發現反應槽移動速率為0.006m/s時能以最快的時間,達到所需的溫度,接著探討熱循環裝置在不同條件下進行反應槽經過熱循環實驗的結果,發現減少反應槽與加熱器間的接觸熱阻,可有效使反應槽的溫度上升,加熱裝置的絕熱能使熱循環時的溫度曲線更穩定,接著以不同體積的反應槽進行熱循環實驗,發現體積較小的反應槽升降溫速率較快。比較以市售PID 溫度控制器和自製單晶片溫控裝置加熱鋁塊進行熱循環的溫度曲線,發現兩者的溫度曲線差異不大,這是因為當加熱溫區皆能維持穩態時,反應槽的升降溫和移動速率有較大的關聯性,因此熱循環裝置只要能精準的控制到所需溫度並維持在穩態溫度即可。最後驗證此熱循環機構的穩定度,結果發現經長時間加熱後反應槽升降溫曲線和初始的反應槽升降溫曲線相似,可證明此熱循環裝置能使溫區溫度維持一致,不受長時間加熱所影響,因此可運用此熱循環裝置投入於長時間加熱之實驗中,如PCR 等生化反應。
In this work we develop an Oscillatory Thermal Cycler Chamber that is combined with the 8051 PWM controller, Smart Motor, three aluminum heaters and chamber. To set the velocity of chamber when chamber move through the Thermal Cycler, we use the simulation software CFDRC to find the optimize parameters. We utilize the user subroutines compiled by FORTRAN language to make the numerical results realistically. We find that the effects of various moving velocities of the chamber on the temperature
distributions can be negligible. The velocity of chamber is set to be 0.0006m/s, and three heating face can make the chamber achieve the temperature which we need.
We investigate the temperature of the chamber which moves through thermal cycler in different situations. Reducing the contact thermal resistance can make temperature of the chamber higher; the heat isolation can make the temperature of chamber more stable. Then we carry on the experiment by using the different volumes of chambers, the heating and the cooling rate of the small chamber is faster than the big chamber. We find that using the heater controlled by 8051 and that by commercial PID controller to III heat the aluminum block, the temperature curves of the chamber are similar.
The temperatures of three aluminum heaters are at steady state, the heating and cooling rates of the chamber are relative to the traveling speed of the chamber, it means the thermal cycler just needs to be maintained at the steady state temperature of aluminum block in. Finally we confirm the stability of this oscillatory thermal cycler chamber, the temperature curve of the chamber at the first cycle is similar to that at the specific cycle after one hour. Therefore we may utilize this oscillatory thermal cycler chamber to perform the PCR experiment in the future.
目 錄
摘 要 ·········································································································· I
Abstract ··········································································································· II
誌 謝 ······································································································· IV
目 錄 ········································································································ V
表 目 錄 ······································································································· IX
圖 目 錄 ········································································································ X
第1 章 緒論 ··································································································· 1
1.1 研究背景 ............................................................................................ 1
1.2 研究動機 ............................................................................................ 2
第2 章 文獻探討 ··························································································· 4
2.1 PCR 晶片 ............................................................................................. 4
2.1.1 微腔體式PCR 晶片 ······························································ 4
2.1.2 連續流體式PCR 晶片 ·························································· 7
2.2 連續流體式PCR 系統 ..................................................................... 14
2.2.1 單一方向式(Unidirectional)CFPCR 系統 ······················ 14
2.2.2 封閉迴路式(Closed Loop)CFPCR 系統 ························· 15
2.2.3 往復式(Oscillatory)CFPCR 系統 ·································· 17
2.3 PCR 裝置與其它次系統整合 ........................................................ 18
VI
2.4. 研究目的 ......................................................................................... 20
第3 章 理論分析與數值方法 ····································································· 22
3.1 理論模式 .......................................................................................... 22
3.2 數值軟體 ........................................................................................... 23
3.3 數值方法 .......................................................................................... 24
3.3.1 網格結構 ·············································································· 24
3.3.2 計算用之統御方程式 ························································· 24
3.3.3 上風算則 ············································································· 25
3.3.4 殘值 ····················································································· 25
3.3.5 鬆弛係數 ·············································································· 26
3.4 使用者副程式 .................................................................................. 26
3.4.1 副程式的範例 ······································································ 26
3.4.2 副程式的建立 ······································································ 29
3.4.3 模擬使用的副程式 ······························································ 29
第4 章 實驗設備與方法 ············································································· 31
4.1 硬體實驗設備 .................................................................................. 31
4.1.1 反應槽 ·················································································· 31
4.1.2 溫度量測系統的製備 ·························································· 33
4.1.3 加熱系統的製備 ·································································· 36
VII
4.1.4 反應槽驅動系統 ································································· 39
4.2 往復式熱循環反應器的製作 ........................................................... 42
4.3 控制軟體 ........................................................................................... 43
4.4 量測方法 ........................................................................................... 48
第5 章 結果與討論 ····················································································· 50
5.1 模擬反應槽在不同加熱面積下於系統內移動 .............................. 50
5.1.1 改變加熱面積 ····································································· 57
5.2 以市售PID 加熱控制器加熱鋁塊 .................................................. 59
5.2.1 反應槽於加熱機構內往復移動之結果······························ 62
5.2.2 絕熱材料和導熱膏的影響 ················································· 63
5.2.3 減少加熱塊厚度,增加空氣對流溫區距離 ······················ 65
5.3 8051 溫控裝置加熱鋁塊 .................................................................. 67
5.3.1 PID 溫控裝置和8051 溫控裝置的比較 ····························· 70
5.3.2 改變反應槽厚度 ································································· 72
5.3.3 8051 溫控裝置的穩定度 ·················································· 73
第6 章 結論與建議 ····················································································· 75
6.1 結論 .................................................................................................. 75
6.2 建議 .................................................................................................. 77
參考文獻 ······································································································· 79
VIII
附錄A.邊界條件副程式 ·············································································· 85
作者簡介 ······································································································· 90
[1] Northrup, M. A., Ching, M. T., White, R. M., and Wltson, R. T., “DNA
Amplification in a Microfabricated Reaction Chamber,” Proceeding of 7th
International Solid-State Sensors and Actuators Conference, Transducers ’93,
pp. 924-926, 1993.
[2] El-Ali, J., Perch-Nielsen, I. R., Poulsen, C. R., Bang, D. D., Telleman, P.,
and Wolff, A., 2004, “Simulation And Experimental Validation of a SU-8
Based PCR Thermocycler Chip with Integrated Heaters And Temperature
Sensor,” Sensors and Actuators A: Physical, Vol. 110, pp. 3-10.
[3] Shen, K., H., Chen, X., Guo, M., Cheng, J., 2005, “A microchip-based
PCR device using flexible printed circuit technology,” Proc of Sensors and
Actuators , Vol. 105, pp. 251-258.
[4] Neuzil, P., Zhang C., Pipper J., Oh S., and Zhuo L., 2006, “Ultra fast
miniaturized real-time PCR: 40 cycles in less than six minutes,” Nucleic Acids
Research, Vol. 34, pp.1-9.
[5] Singh, J., and Ekaputri, M., 2006, “PCR thermal management in an
integrated Lab on Chip,” International MEMS Conference, Series.34, pp.
222-227.
[6] Dahl, A., Sultan, M., Jung, A., Schwartz, R., Lange, M., Steinwand, M.,
Livak, K., J., Lehrach, H., Nyarsik, L., 2007, “Quantitative PCR based
expression analysis on a nanoliter scale using polymer nano-well chips,”
Biomed Microdevices, Vol. 9, pp. 307-314.
[7] Liua, H., B., Ramalingam, N., Jiang, Y., Dai, C., C., Hui, K., M., Gong, H.,
Q., 2008, “Rapid distribution of a liquid column into a matrix of nanoliter
wells for parallel real-time quantitative PCR,” Sensors and Actuators B:
Chemical, Vol. 4982, pp. 1-7.
[8] Kopp, M. U., A. Mello de, J. and Manz, A., 1998, “Chemical
80
Amplification: Continuous-Flow PCR on a Chip,” Science, Vol. 280, No.
5366, pp. 1046-1048.
[9] Schneega, I., Brautigamb, R., and Kohlera, J., M., 2001, “Miniaturized
flow-through PCR with different template types in a silicon chip
thermocycler,” Lab on a Chip, Vol.1. pp. 42-49.
[10] Zhang, Q., Wang, W., Zhang, H., and Wang, Y., 2002, “Temperature
Analysis of Continuous-Flow Micro-PCR Based on FEA,” Sensors and
Actuators B: Chemical, Vol. 82, No.1, pp. 75-81.
[11] Sugumar, D., Ashraf, M. A., and KongL, X., 2006, “Computational
Thermal Analysis of a Continuous Flow Micro Polymerase Chain Reaction
(PCR) Chip,” Proceeding of SPIE, Vol. 6415, pp. 641517-1-641517-7.
[12] Shih, C., Y., Chen, Y., Tai, Y., C., 2006, “Parylene-strengthened thermal
isolation technology for microfluidic system-on-chip applications,” Sensors
and Actuators A, Vol. 126, pp. 270-276.
[13] Joung, S. R., Kang, C. J., and Kim, Y. S., 2008, “Series DNA
Amplification Using the Continuous-Flow Polymerase Chain Reaction Chip,”
Japanese Journal of Applied Physic, Vol. 47, pp. 1342-1345.
[14] Crews, N., Wittwer, C., and Gale, B., 2008, “Continuous-flow thermal
gradient PCR,” Biomed Microdevices, Vol.10, pp.187-195.
[15] Sun, K., Yamaguchi, A., Ishida, Y., Matsuo, S., and Misawa, H., 2002, “A
Heater-Integrated Transparent Microchannel Chip for Continuous-Flow
PCR,” Sensors and Actuators B: Chemical, Vol. 82, pp. 283-289.
[16] Nakayama, T., Kurosawa, Furui, Y, Kerman, S., Kobayashi, K., Rao, S.
R., Yonezawa Y., Nakano, K., Hino, A., Yamamura, S., Takamura, Y., and
Tamiya, E., 2006, “Circumventing Air Bubbles in Microfluidic Systems And
Quantitative Continuous-Flow PCR Applications,” Analytical and
Bioanalytical Chemistry, Vol. 386, No. 5, pp. 1327-1333.
81
[17] Yu, C., Liang, W., Kuan, I., Wei, C., and Gu, W., 2007, “Fabrication And
Characterization of a Flow-Through PCR Device with Integrated Chromium
Resistive Heaters” Journal of the Chinese Institute of Chemical
Engineers ,Vol. 38,No. 3-4, pp. 333-339.
[18] Obeid, P. J., Christopoulos, T. K., Crabtree, H. J., and Backhouse, C. J.,
2003, “Microfabricated Device for DNA And RNA Amplification by
Continuous-Flow Polymerase Chain Reaction And Reverse
Transcription-Polymerase Chain Reaction with Cycle Number Selection,”
Analytical Chemistry, Vol. 75, No. 2, pp. 288-295.
[19] Mitchell, M., W., Liu, X., Bejat, Y., Nikitopoulos, D., E., 2003,
“Modeling and Validation of a Molded Polycarbonate Continuous Flow
Polymerase Chain Reaction Device,” Proceedings of SPIE, Vol. 4982, pp.
83-98.
[20] Hashimoto, M., Chen, P. C., Mitchell, M. W., Nikitopoulos, D. E., Soper,
S. A., and Murphy, M. C., 2004, “Rapid PCR in a Continuous Flow Device,”
Lab on a Chip, Vol. 4, No.4, pp. 638-645.
[21] Xiaoyu, J., Zhiqiang, N., Wenyuan, C., and Weiping, Z., 2005,
“Polydimethylsiloxane (PDMS)-Based Spiral Channel PCR Chip,”
Electronics Letters, Vol. 41, No.16, pp. 11-12.
[22] Chen, P. C., Nikitopoulos, D. E., Soper, S. A., and Murphy, M. C., 2008,
“Temperature Distribution Effects on Micro-CFPCR Performance,”
Biomedical Microdevices, Vol. 10, No. 2, pp. 141-152.
[23] Chou, C. F., Changrani, R., Roberts, P., Sadler, D., Burdon, J.,
Zenhausern, F., Lin, S., Mulholland, A., Swami, N., and Terbrueggen, R.,
2002, “A Miniaturized Cyclic PCR Device-Modeling And Experiments,”
Microelectronic Engineering, Vol. 61-62, No. 27, pp. 921-925.
[24] West, J., Karamata, B., Lillis, B., Gleeson, J. P., Alderman, J., Collins, J.
K., Lane, W., Mathewson, A., and Berney, H., 2002, “Application of
Magnetohydrodynamic Actuation to Continuous Flow Chemistry,” Lab on a
82
Chip, Vol. 2, No. 4, pp. 224-230.
[25] Liu, J., Enzelberger, M., and Quake, S., 2002, “A Nanoliter Rotary
Device for Polymerase Chain Reaction,” Electrophoresis, Vol. 23, No. 10, pp.
1531-1536.
[26] Sun, Y., Nguyen, N. T., and Kwok, Y. C., 2008, “High-Throughput
Polymerase Chain Reaction in Parallel Circular Loops Using Magnetic
Actuation,” Analytical Chemistry, Vol. 80, No. 15, pp. 6127-6130.
[27] Bu, M., Melvin, T., Ensell, G., Wilkinson, J. S., and Evans, A. G. R., 2003,
“Design And Theoretical Evaluation of a Novel Microfluidic Device to Be
Used for PCR,” Journal of Micromechanics and Microengineering, Vol. 13,
pp. 125-130.
[28] Wang, W., Li, Z. X., Luo, R., Lu, S. H., Xu, A. D., and Yang, Y. J., 2005,
“Droplet-Based Micro Oscillating-Flow PCR Chip,” Journal of
Micromechanics and Microengineering, Vol. 15, pp. 1369-1377.
[29] Hua, G., Xiang, Q., Fu, R., Xu, B., Venditti, R., Li, D., 2006,
“Electrokinetically controlled real-time polymerase chain reaction in
microchannel using Joule heating effect,” Analytica Chimica Acta, Vol. 557,
pp. 146-151.
[30] Curcio, M., and Roeraade, J., 2003, “Continuous Segmented-Flow
Polymerase Chain Reaction for High-Throughput Miniaturized DNA
Amplification,” Analytical Chemistry, Vol. 75, No. 1, pp. 1-7.
[31] Park, N., Kim, S., and Hahn, J. H., 2003, “Cylindrical Compact
Thermal-Cycling Device for Continuous-Flow Polymerase Chain Reaction,”
Analytical Chemistry, Vol. 75, No. 21, pp. 6029-6033.
[32] Chabert, M., Dorfman, K., D., Cremoux, P., d., Roeraade, J., and Viovy,
J., L., 2006, “Automated Microdroplet Platform for Sample Manipulation and
Polymerase Chain Reaction,” Analytical Chemistry, Vol. 78, No. 22, pp.
7722-7728.
83
[33] Chiou, J., Matsudaira, P., Sonin, A., and Ehrlich, D., 2001, “A
Closed-Cycle Capillary Polymerase Chain Reaction Machine,” Analytical
Chemistry, Vol. 73, No. 9, pp. 2018-2021
[34] Chen, Z., Qian, S., Abrams, W. R., Malamud, D., and Bau, H. H., 2004,
“Thermosiphon-Based PCR Reactor: Experiment and Modeling,” Analytical
Chemistry, Vol. 76, No. 13, pp. 3707-3715.
[35] Zhang, C., Xing, D., 2009, “Parallel DNA amplification by convective
polymerase chain reaction with various annealing temperatures on a thermal
gradient device,” Analytical Biochemistry, Vol. 387, pp.102-112.
[36] Chen, L., West, J., Auroux, P. A., Manz, A., and Day, P. J. R., 2007,
“Ultrasensitive PCR And Real-Time Detection from Human Genomic
Samples Using a Bidirectional Flow Microreactor,” Analytical Chemistry, Vol.
79, No. 23, pp. 9185-9190.
[37] Frey, O., Bonneick, S., Hierlemann, A., and Lichtenberg, J., 2007,
“Autonomous microfluidic multi-channel chip for real-time PCR with
integrated liquid handling,” Biomed Microdevices, Vol.7, pp.711-718.
[38] Tsuchiya, H., Okochi, M., Nagaob, N., Shikida, M., Honda, H., 2008,
“On-chip polymerase chain reaction microdevice employing a magnetic
droplet-manipulation system,” Sensors and Actuators, Vol.30, pp. 583-588.
[39] Bahrami, M., Melvin, T., Wilkinson, J. S., and Evans, A. G. R., 2005,
“PCR Device with Integrated Thermal Cycling and Fluorescence Detection
Elements,” Proceedings of SPIE, Vol. 5836, No. 2, pp. 285-292.
[40] Gartner, C., Klemm, R., and Holger, B., 2007, “Methods And
Instruments for Continuous-Flow PCR on a Chip” Proceedings of SPIE, Vol.
6465, No. 646502, pp. 646502-1-646502-8.
[41] Tsai, N. C., and Sue, C. Y., 2007, “Thermal Control of Micro Reverse
Transcription-Polymerase Chain Reaction Systems,” Sensors and Actuators A:
Physical, Vol. 136, No. 1, pp. 178-183.
84
[42] Hsieh, T. M., Luo, C. H., Huang, F. C., Wang, J. H., Chien, L. J., and Lee,
G. B., 2008, ”Enhancement of Thermal Uniformity for a Microthermal Cycler
And Its Application for Polymerase Chain Reaction,” Sensors And Actuators
B: Chemical, Vol. 130, No. 2, pp. 848-856.
[43] Lee, D. S., and Chen, C. S., 2008, “Development of a Temperature
Sensor Array Chip And a Chip-Based Real-Time PCR Machine for DNA
Amplification Efficiency-Based Quantification,” Biosensors and
Bioelectronics, Vol. 23, No.7, pp. 971-979.
[44] Sudip, M., and Venkataraman, V., 2007, “Novel Fluorescence Detection
Technique for Non-Contact Temperature Sensing in Mcrochip PCR,”
Proceeding of Biochemical and Biophysical Methods, Vol.70, pp. 773-777.
[45] Kim, S., H., Noh, J., Jeon, M., K., Kim, K., W., Lee, L., P., and Woo, S.,
I., 2006 “Micro-Raman thermometry for measuring the temperature
distribution inside the microchannel of a polymerase chain reaction chip,”
Proc of Micromechanics and Microengineering, Vol. 16, pp. 526-530.
[46] Korampally, V., Bhattacharya, S., Gao, Y., Grant, S. A., Kleiboeker, S. B.,
Gangopadhyay, K., Tan, J., and Gangopadhyay, S., 2006, “Optimization of
Fabrication Process for a PDMS-SOG-Silicon Based PCR Micro Chip
through System Identification Techniques,” Proceeding of 19th IEEE
Symposium on Computer-Based Medical Systems, pp. 329-334.
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