On Demand Release and Retrieval of Bacteria from Microwell Arrays Using Photodegradable Hydrogel Membranes
On Demand Release and Retrieval of Bacteria from Microwell Arrays Using Photodegradable Hydrogel Membranes
复制标题
使用可光降解水凝胶膜从微孔阵列中按需释放和回收细菌
DOI：
10.1021/acsabm.8b00592
复制
发表时间：
2018
影响因子：
4.7
通讯作者：
Hansen, Ryan R.
中科院分区：
文献类型：
--
作者：
van der Vlies, André J.;Barua, Niloy;Nieves-Otero, Priscila A.;Platt, Thomas G.;Hansen, Ryan R.
微孔阵列是研究微生物和哺乳动物系统中单细胞行为和细胞间相互作用的重要工具。然而，从高空间精度的微孔阵列中提取细胞仍然是一个主要的技术障碍，阻碍了观察到的微孔中细胞的后续遗传和表型表征。这项工作描述了一种新的、基于材料的方法，利用植物病原体农杆菌作为模型细菌，在阵列中从小（直径≥20 μm）微孔中生长和回收活细菌细胞。我们的方法使用光响应，步聚聚（乙二醇）水凝胶界面作为膜，将活动细胞限制在微孔内，同时允许营养交换和细胞生长。关键的设计特点是膜的光降解性，因为它可以使用图案紫外光打开感兴趣的单个井，以选择性地释放和回收细胞。可以从用户定义的任意数量和组合的井中并行抽取。这些进步代表了光响应水凝胶的新用途，并且能够以高空间精度从微孔中检索细胞，这使得需要从异质群体中分离和表征具有罕见表型的细胞的几种应用成为可能。
Microwell arrays are important tools for studying single cell behavior and cell–cell interactions, both in microbial and mammalian systems. However, retrieval of cells from microwell arrays with high spatial precision remains a major technical hurdle that prevents follow-up genetic and phenotypic characterization of cells within observed microwells. This work describes a new, material-based approach to grow and retrieve live bacterial cells from small (≥20 μm diameter) microwells in an array using the plant pathogenAgrobacterium tumefaciensas a model bacterium. Our approach uses a light-responsive, step-polymerized poly(ethylene glycol) hydrogel interface as a membrane that confines motile cells within microwells while allowing nutrient exchange and cell growth. The key design feature is the photodegradability of the membrane, as it enables individual wells of interest to be opened using patterned UV light for selective release and retrieval of cells. Extraction can occur in parallel from any number and combination of wells defined by the user. These advancements represent a new use for light-responsive hydrogels and the ability to retrieve cells from microwells with high spatial precision enables several applications that require the isolation and characterization of cells with rare phenotypes from heterogeneous populations.