2002 Fiscal Year Final Research Report Summary
Development and application of a novel capillary chromatography system utilizing vital reaction of a single cell
| Project/Area Number |
13480294
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Osaka University |
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Principal Investigator |
ARAKI Tsutomu Osaka Univ., Engineering Science, Prof., 基礎工学研究科, 教授 (50136214)
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| Co-Investigator(Kenkyū-buntansha) |
TOHNO Yoshiyuki Nara Medical University. Prof., 医学部, 教授 (40075023)
YASUI Takeshi Osaka Univ., Engineering Science, Res. Assist., 基礎工学研究科, 助手 (70314408)
HASHIMOTO Mamoru Osaka Univ., Engg. Science, Assoc. Prof., 基礎工学研究科, 助教授 (70237949)
CHIKAMORI Kensuke Naruto Univ.of Education, Assos. Prof., 学校教育学部, 助教授 (40108874)
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| Project Period (FY) |
2001 – 2002
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| Keywords | MICROSENSOR / CELL SENSOR / CAPILLARY ELECTROPHORESIS / SACCHAROMYCES CEREVISIAE / FLUORESCENCE MICROSCOPE / SECOND HARMONIC GENERATION LIGHT / PARAMECIUM CAUDATUM / FEMTOSECOND LASER |
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| Research Abstract |
We have proposed a biosensor chromatography system in conjunction with a capillary electrophoresis (CE) chromatography and a living cell to monitor the cellular environment. As the sensor cell for bio-CE system, Saccharomyces cerevisiae (SC) was used, SC was stained with rhodamine 123 (Rhl23) that is sensitive to electric potential of the mitochondria membrane. The cell was enveloped inside the capillary, and resultant Rh123 fluorescence image was measured with a fluorescence microscope and a CCD system.
In order to avoid unwanted electric field of 5 kV to the sensor cell, a micro-crack that works as the electric by-pass is attractive. To make the by-pass, micro-processing by a femtosecond (fs) laser was applied We have achieved to hole the glass wall of the capillary tube with a few micrometer in diameter, however the processing has not completed yet because of lack of the precision for the laser beam focusing inside the small wall. Further trial is required to succeed in the production of the micro hole. When the pulsed light from the fs laser is irradiated to the biological tissue, second harmonic generation light (SHG light) is induced. Based on this phenomenon, we have constructed a polarization SHG light detection system. We found that orientation of the collagen fiber has been informed by the SHG light.
As another trial of the cell sensor system, we have utilized a Paramecium caudatum as the sensor cell to monitor the water pollution based on the analysis of the cell moving profile in a micro-chamber. The paramecium cell showed characteristic moving profiles due to the pollutant such as heavy metal ions and acid. We have also studied the emission characteristics of a white light emitting diode to evaluate applicability to the compact light source in the present sensor system.
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