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Development and Optimization of a Process for Automated Recovery of Single Cells Identified by Microengraving

机译:通过微雕刻确定的单细胞自动回收方法的开发和优化

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Microfabricated devices are useful tools for manipulating and interrogating large numbers of single cells in a rapid and cost-effective manner, but connecting these systems to the existing platforms used in routine high-throughput screening of libraries of cells remains challenging. Methods to sort individual cells of interest from custom microscale devices to standardized culture dishes in an efficient and automated manner without affecting the viability of the cells are critical. Combining a commercially available instrument for colony picking (CellCelector, AVISO GmbH) and a customized software module, we have established an optimized process for the automated retrieval of individual antibody-producing cells, secreting desirable antibodies, from dense arrays of subnanoliter containers. The selection of cells for retrieval is guided by data obtained from a high-throughput, single-cell screening method called microengraving. Using this system, 100 clones from a mixed population of two cell lines secreting different antibodies (12CA5 and HYB099-01) were sorted with 100% accuracy (50 clones of each) in ~2 h, and the cells retained viability.
机译:微型设备是用于以快速且经济高效的方式操纵和询问大量单细胞的有用工具,但是将这些系统连接到用于细胞库常规高通量筛选的现有平台仍然具有挑战性。在不影响细胞生存力的前提下,以有效和自动化的方式将感兴趣的单个细胞从定制的微型设备分类到标准化培养皿的方法至关重要。结合市售的菌落拾取工具(CellCelector,AVISO GmbH)和定制的软件模块,我们建立了一种优化的过程,可从致密的亚纳升容器阵列中自动检索分泌所需抗体的单个抗体生产细胞。从取自高通量单细胞筛选方法(称为微雕刻)的数据中选择要检索的细胞。使用该系统,在约2小时内以100%的准确度(来自50个克隆)对来自分泌不同抗体(12CA5和HYB099-01)的两种细胞系混合种群的100个克隆进行了分选,并且细胞保持了活力。

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