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.

我们已经提出了与毛细管电泳（CE）色谱和活细胞一起监测细胞环境的生物传感器色谱系统。作为生物CE系统的传感器细胞，使用了酿酒酵母（SC），将SC用若丹明123（RHL23）染色，对线粒体膜的电势敏感。将细胞包裹在毛细管内，并用荧光显微镜和CCD系统测量所得的RH123荧光图像。为了避免对传感器电池的不需要的5 kV的电场，这是一种可作为电气旁poss的微型裂缝，是有吸引力的。为了使旁路，使用飞秒（FS）激光器进行微处理，我们已经实现了用直径为几微米的毛细管的玻璃壁，但是由于缺乏在小壁内部的激光束聚焦的精确度，因此该处理尚未完成。需要进一步的试验才能成功生产微孔。当将FS激光器的脉冲光照射到生物组织中时，诱发了第二次谐波产光（SHG光）。基于这种现象，我们构建了一个极化SHG光检测系统。我们发现，胶原蛋白纤维的方向已通过SHG光告知。在细胞传感器系统的另一项试验中，我们利用了caudatum caudatum作为传感器细胞来监测水污染，基于对微型室中细胞移动谱的分析。由于污染物（例如重金属离子和酸），甲状腺菌细胞显示出特征性移动曲线。我们还研究了发射二极管的发射特性，以评估当前传感器系统中紧凑型光源的适用性。