Ran TU1(), Shixin LI2, Haoni LI2, Meng WANG1
Received:
2021-12-03
Contact:
Meng WANG
E-mail:tu_r@tib.cas.cn
CLC Number:
Ran TU, Shixin LI, Haoni LI, Meng WANG. Advances and applications of droplet-based microfluidics in evolution and screening of engineered microbial strains[J]. Synthetic Biology Journal, doi: 10.12211/2096-8280.2021-105.
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URL: https://synbioj.cip.com.cn/EN/10.12211/2096-8280.2021-105
Tab. 1
Droplet-based screening system and application using FADS
液滴类型 | 液滴信号来源 | 优缺点 | 通量 | 代表性应用案例 |
---|---|---|---|---|
单层液滴 | 细胞自身信号(荧光、拉曼、浊度、细胞图像) | 优点:操作简单,细胞直接包埋在液滴中。 缺点:需要特殊软件,例如拉曼和图像识别软件。 | 荧光300 Hz;拉曼1~5 Hz;图像识别5~10 Hz;浊度240 Hz | 色素:核黄素[ |
细胞荧光蛋白(目标蛋白和外源荧光蛋白融合、生产菌自身含生物传感器、生产菌和感应菌共包埋培养) | 优点:操作简单,细胞直接包埋在液滴中。 缺点:需要构建特定生物传感器。 | 150~300 Hz | 生产菌自身含生物传感器:3-脱氢莽草酸[107;生产菌和感应菌共培养:香豆酸酵母菌和大肠杆菌感应报告菌[ | |
细胞和底物(同时共包埋在液滴) | 优点:操作简单,细胞和底物直接包埋在液滴中。 缺点:不适合酶促反应快的样品。 | 300~2000 Hz | 大肠杆菌:半乳糖苷酶[ | |
细胞和底物(细胞先包埋在液滴,底物后加入) | 优点:可以通过液滴融合添加试剂,用于酶促反应快的样品。 缺点:液滴融合操作复杂。 | 300~1000 Hz | 漆酶[ |
Tab. 2
Droplet-based screening system and application using FACS
液滴类型 | 液滴信号来源 | 优缺点 | 通量 | 代表性应用案例 |
---|---|---|---|---|
双层液滴 | 细胞和底物(同时共包埋在液滴)、细胞代谢物自发荧光等 | 优点:液滴无需固化处理。 缺点:制备双层液滴操作复杂;无法进行液滴融合添加其他试剂。 | 3~20 kHz | 酯酶[ |
凝胶液滴 | 细胞荧光蛋白(生产菌自身含生物传感器、生产菌和感应菌共包埋培养)、细胞和底物(同时共包埋在液滴) | 优点:操作简单,直接制备凝胶液滴。 缺点:凝胶液滴需固化处理;无法进行液滴融合添加其他试剂。 | 1~6 kHz(FACS分选仪);30~40 Hz (COPAS大颗粒分选仪) | 脂肪酶[ |
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