2012 Fiscal Year Final Research Report
Real-time and on-site monitoring of bacterial cells in freshwater environments by using a microfluidic system
| Project/Area Number |
21256002
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 海外学術 |
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| Research Field |
Environmental pharmacy
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| Research Institution | Osaka University |
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Principal Investigator |
NASU Masao 大阪大学, 大学院・薬学研究科, 教授 (90218040)
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| Co-Investigator(Kenkyū-buntansha) |
TANI Katsuji 大阪大谷大学, 薬学部, 教授 (50217113)
KAWAI Mako 姫路獨協大学, 薬学部, 准教授 (40533922)
YAMAGUCHI Nobuyasu 大阪大学, 大学院・薬学研究科, 准教授 (20252702)
BABA Takashi 大阪大学, 大学院・薬学研究科, 助教 (20423121)
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| Co-Investigator(Renkei-kenkyūsha) |
KAWABATA Zen'ichiro 総合地球環境学研究所, 研究部, 教授 (80108456)
IWAMOTO Tomotada 神戸市環境保健研究所, 微生物部, 副部長 (70416402)
ICHIJO Tomoaki 大阪大学, 大学院・薬学研究科, 特任研究員 (20513899)
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| Research Collaborator |
陳 功祥 中国, 浙江大学医学部, 教授
HIEN Lai Thuy ベトナム, 科学技術アカデミー, 准教授
MUANGKAEW Kanya タイ, 科学技術省, 研究員
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| Project Period (FY) |
2009 – 2012
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| Keywords | 水環境 / on-site モニタリング / 細菌 / アジア / マイクロ流路デバイス |
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| Research Abstract |
“Real-time” and “on-site” microbiological methods are required for prevention of waterborne diseases. We fabricated a portable microfluidic system (36 cm × 54 cm × 23 cm, 15 kg) for rapid monitoring of bacterial cells in freshwater at a density in the order of 104- 106/ml.A microfluidic device (size: 48 mm × 23 mm) was designed and fabricated using polydimethylsiloxane (PDMS) and a thin glass sheet. Escherichia coli O157:H7 and Legionella pneumophila cells were spiked in purified freshwater (101- 106cells/ml). Low numbers of targeted cells were collected on a filter and then resuspended in purified freshwater for a 100 to 1,000 -fold concentration. Fluorescent dye was used for nucleic acid stain and direct counting of total bacterial cells. Fluorescent antibody was used for specific detection of E. coli O157:H7 and L. pneumophila cells in the samples. The portable microfluidic system was used for the detection of fluorescently stained cells flowing i n the microchannel, and number of the cells in each sample was determined using original image analysis software. Bacterial cells in the same samples were counted by fluorescence microscopy for comparison. The numbers of bacterial cells determined using the microfluidic system was highly correlated with the microscopic counts. We could count targeted cells in the samples within 2 hours with this microfluidic system. The microfluidic device and portable counting system fabricated in this study will contribute to the microbiological quality control of freshwater.
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