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目前主要使用激光共聚焦扫描显微镜观察绿色荧光蛋白的表达,但需要昂贵的仪器并耗费大量时间。本研究开发了一种新型激光诱导的微流芯片检测系统来监测绿色荧光蛋白在枯草芽孢杆菌中的表达。该系统主要由激光装置、光路系统、微流控芯片、光电倍增管和计算机处理系统等5部分组成。对该系统的测试结果显示,随着诱导强度的增强监测信号峰也随之增强,并且与激光共聚焦显微镜观察的结果一致。利用该芯片系统能够快速准确地筛选和鉴定用绿色荧光蛋白作为标记的细胞克隆,可以替代PCR鉴定方法。但该系统仅仅能够监测表达强度,不能够满足蛋白定位等高水平研究,因此,该系统适合应用于环境的微生物监测、药物筛选和其他无需观察蛋白定位等研究。
The current use of laser scanning confocal microscopy of green fluorescent protein expression, but requires expensive equipment and consume a lot of time. This study developed a new laser-induced microfluidic chip detection system to monitor the expression of green fluorescent protein in Bacillus subtilis. The system is mainly composed of 5 parts such as laser device, optical system, microfluidic chip, photomultiplier tube and computer processing system. The test results of this system showed that the monitoring signal peak also increased with the enhancement of the induction intensity, and it was consistent with the result of laser confocal microscopy. The chip system can rapidly and accurately screen and identify the green fluorescent protein as a marker cell clone, which can replace the PCR identification method. However, the system can only monitor the expression intensity and can not meet the high level of protein localization. Therefore, the system is suitable for microbial monitoring in the environment, drug screening and other studies that do not need to observe the protein localization.