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Development and application of a novel capillary chromatography system utilizing vital reaction of a single cell

利用单细胞生命反应的新型毛细管色谱系统的开发与应用

基本信息

批准号：
13480294
负责人：
ARAKI Tsutomu
金额：
$ 9.41万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

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.

我们提出了一种生物传感器色谱系统，结合毛细管电泳（CE）色谱和活细胞来监测细胞环境。以酿酒酵母（Saccharomycescerevisiae，SC）为传感细胞，用对线粒体膜电位敏感的罗丹明123（Rhl23）对SC进行染色。为了避免5 kV的电场对传感器细胞的干扰，在传感器表面设计了一个微裂纹作为电旁路，并利用荧光显微镜和CCD系统测量了Rh123荧光图像。为了制造旁路，应用了飞秒（fs）激光的微加工我们已经实现了在直径几微米的毛细管玻璃壁上打孔，但由于激光束聚焦精度不够，加工尚未完成小壁内。需要进一步的试验才能成功地生产微孔。当来自fs激光器的脉冲光照射到生物组织时，感应出二次谐波产生光（SHG光）。基于这一现象，我们构建了一个偏振倍频光探测系统。作为细胞传感器系统的另一个尝试，我们利用草履虫作为传感细胞，通过对细胞在微腔中运动轮廓的分析，对水体污染进行了监测。草履虫细胞在重金属离子和酸等污染物的作用下呈现出特征性的运动轮廓。我们还研究了白色发光二极管的发射特性，以评估在本传感器系统中对紧凑光源的适用性。