Development of micro in situ analysis devices for microbiology in oceanic environment

海洋环境微生物微型原位分析装置研制

• 批准号: 14205147
• 负责人: YAMAMOTO Takatoki
• 金额: $ 34.69万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14205147/
• 关键词: microfluidic device; in situ analysis system; gene analysis; deep-sea; IISA-Gene; PCR; Microbes; PDMS; 現場型装置; マイクロデバイス; DNA精製; 集積化; 現場型遺伝子解析装置; マイクロ加工技術; micro TAS; PDMS(Polydimethylsiloxane); Flow-through PCR; 極限環境; マイクロ分析チップ; 現場型分析装置

In this research, an in situ analysis device for microbiology is developed and evaluated with microfabrication techniques. As a result, IISA (Integrated In situ Analyzer) -Gene was successfully developed with compact format. To achieve the device, development and evaluation was performed as follows :(1)A PDMS based microchannel chip and a glass based temperature control chip was fabricated to perform molecular analysis of microbes living in oceanic environments with PCR method.(2)Surfactant reagent (Tween 20) was applied to prevent protein adsorption onto PDMS surface. It increased reaction performance effectively.(3)Chaotropic reagent (GuSCN) and glass beads based DNA purification function was integrated to the device.(4)LED light source and photo detector (PMT) was integrated to the device for optical detection of PCR products.(5)For in situ operation, miniaturized pumping system with pumps and valves is connected to the microchip.(6)Microprocessor based controller circuit and power supply are packaged in a pressure resistant housing. IISA-Gene was mounted in oil-filled housing for pressure equalization.(7)High-pressure experiment system was developed and utilized for evaluations of the IISA-Gene. It can simulate high pressure (up to 60 MPa) and low temperature (down to 2℃).The IISA-Gene was operated in real deep-sea environment with RV "NATSUSHIMA" and ROV "HYPERDOLPHIN" (JAMSTEC, Japan) at off Ishigaki Isl. Japan. As a result, the system was operated successfully and the primitive data was obtained.

本研究开发了一种微生物学原位分析装置，并利用微细加工技术对其进行了评价。因此，IISA(集成原位分析仪)-基因的开发成功，格式紧凑。(1)制作了基于PDMS的微通道芯片和基于玻璃的温控芯片，用于海洋环境微生物的分子分析。(2)应用表面活性剂(Tween20)阻止蛋白质在PDMS表面的吸附。该装置集成了基于杂化试剂(GuSCN)和玻璃微珠的DNA纯化功能。(4)将LED光源和光电探测器(PMT)集成到装置中，用于对PCR产物进行光学检测。(5)对于现场操作，微型泵送系统与微芯片相连。(6)基于微处理器的控制器电路和电源封装在耐压外壳中。将IISA-gene安装在充满油的壳体中进行压力均衡。(7)研制了IISA-gene高压实验系统，并对其进行了评价。它可以模拟高压(高达60 Mpa)和低温(低至2℃)。IISA-Gen在石垣岛附近的真实深海环境中与RV“Natsushima”和ROV“HYPERDOLPHIN”(日本，JAMSTEC)一起运行。日本。结果，该系统运行成功，获得了原始数据。

项目成果

福場辰洋, 藤井輝夫: "マイクロファブリケーション技術を応用した遺伝子解析システムの開発とその展望"ゲノム医学. 71. 577-584 (2003)

Tatsuhiro Fukuba、Teruo Fujii：“应用微加工技术的基因分析系统的开发及其前景”Genome Medicine 71. 577-584 (2003)。

T.Fukuba, T.Yamamoto, T.Naganuma, T.Fujii: "Microfabricated Flow-through PCR Device for DNA Amplification-Towards in Situ Gene Analysis"Chemical Engineering Journal. (In press). (2004)

T.Fukuba、T.Yamamoto、T.Naganuma、T.Fujii：“用于 DNA 扩增的微型流通式 PCR 装置 - 迈向原位基因分析”化学工程杂志。

Development of an in situ gene analysis device for extreme environmental microbiology

极端环境微生物学原位基因分析装置的研制

• 发表时间: 2005
• 期刊: Oceanography in Japan 14
• 作者: T.Fukuba;T.Yamamoto;T.Naganuma;T.Fujii
• 通讯作者: T.Fujii

Development of miniaturized in situ analysis devices for biological and chemical oceanography

• DOI: 10.1109/mmb.2005.1548383
• 发表时间: 2005-05
• 期刊: 2005 3rd IEEE/EMBS Special Topic Conference on Microtechnology in Medicine and Biology
• 作者: T. Fukuba;A. Imhof;M. Matsunaga;N. Takagi;T. Yamamoto;K. Okamura;T. Naganuma;T. Fujii

Development and field evaluation of an integrated in situ analyzer (IISA) -Gene system

集成原位分析仪（IISA）-基因系统的开发和现场评估

• 发表时间: 2006
• 期刊: Blue Earth '06
• 作者: T.Fukuba;T.Okamoto;T.Naganuma;T.Fujii
• 通讯作者: T.Fujii