Synthetic Biology Journal ›› 2023, Vol. 4 ›› Issue (1): 165-184.DOI: 10.12211/2096-8280.2021-105
• Invited Review • Previous Articles Next Articles
Ran TU1,2, Shixin LI3, Haoni LI3, Meng WANG2
Received:
2021-12-05
Revised:
2022-01-18
Online:
2023-03-07
Published:
2023-02-28
Contact:
Meng WANG
涂然1,2, 李世新3, 李昊霓3, 王猛2
通讯作者:
王猛
作者简介:
基金资助:
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, 2023, 4(1): 165-184.
涂然, 李世新, 李昊霓, 王猛. 液滴微流控技术在微生物工程菌株选育中的应用进展[J]. 合成生物学, 2023, 4(1): 165-184.
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URL: https://synbioj.cip.com.cn/EN/10.12211/2096-8280.2021-105
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 [ | |
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 [ |
Table 1 Droplet-based screening systems and applications of 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 [ | |
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 [ |
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 [ |
Table 2 Droplet-based screening systems and applications of 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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