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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Design and Implementation of an Automated Illuminating Culturing and Sampling System for Microbial Optogenetic Applications
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Design and Implementation of an Automated Illuminating Culturing and Sampling System for Microbial Optogenetic Applications

机译：微生物光遗传学应用自动照明培养和采样系统的设计与实现

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Optogenetic systems utilize genetically-encoded proteins that change conformation in response to specific wavelengths of light to alter cellular processes. There is a need for culturing and measuring systems that incorporate programmed illumination and stimulation of optogenetic systems. We present a protocol for building and using a continuous culturing apparatus to illuminate microbial cells with programmed doses of light, and automatically acquire and analyze images of cells in the effluent. The operation of this apparatus as a chemostat allows the growth rate and the cellular environment to be tightly controlled. The effluent of the continuous cell culture is regularly sampled and the cells are imaged by multi-channel microscopy. The culturing, sampling, imaging, and image analysis are fully automated so that dynamic responses in the fluorescence intensity and cellular morphology of cells sampled from the culture effluent are measured over multiple days without user input. We demonstrate the utility of this culturing apparatus by dynamically inducing protein production in a strain of Saccharomyces cerevisiae engineered with an optogenetic system that activates transcription.
机译：光遗传学系统利用遗传编码的蛋白质来响应特定波长的光而改变构象，从而改变细胞过程。需要结合编程的照明和光遗传学系统的刺激的培养和测量系统。我们提出了一个协议，用于建立和使用连续培养设备以编程剂量的光照射微生物细胞，并自动获取和分析废水中细胞的图像。该设备作为恒化器的操作使得可以严格控制生长速率和细胞环境。定期采样连续细胞培养物的流出物，并通过多通道显微镜对细胞成像。培养，取样，成像和图像分析是完全自动化的，因此无需用户输入，即可在数天内测量从培养液中取样的细胞的荧光强度和细胞形态的动态响应。我们通过动态诱导激活转录的光遗传学系统工程改造的酿酒酵母菌株中诱导蛋白质生产来证明这种培养设备的实用性。

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