Fluorescence-Activated on-Chip Cell Culture Sorting (O3CS): Smart Petri Dish

G MolchanovPavel, E Moreno-CuevasJorge, Hernández-Torre Martín, Gonzalez-Garza Maria Teresa, Garcia Charles, Hillhouse Edward William, L. Christine, M ZsigmondEva, W. Rick, M. John, Zeng Hong, Kim Jeewon, Yang Jieying, Cuevas Raquel, M BulaiPavel, N PitlikTaras, A DenisovAndrey, N CherenkevichSergey, Molchanova Alla Yu, N GolubevaElena, A StrukovVictor, Boksha Victor
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Abstract

On-chip cell sorting is a promising technique for sorting stem cells in culture. On-chip cell sorting allows minimization of lab personnel involvement in cells processing, dramatically reducing the risk of cell culture contamination. We developed a fluorescence-activated On-Chip Cell Culture Sorting (O3CS) system, which combines a biocompatible semiconductor light addressable microarray (chip) and optical setup for chip addressing and cell culture observation. The optical setup has fluorescent and reflected-light microscope capability for visualization and control of cell populations. High-resolution detection of ‘unwanted’ cells with a high-efficient sorter, based on light-induced electroporation is in the core of the O3CS implemented in NeuroSyntek StemOptimizer 6+. We demonstrated capability of the system to perform cell culture fluorescence activated sorting by inducing irreversible single-cell electroporation, validated O3CS sorting efficacy with fluorescent microscopy and flow cytometry, and compared it with the magnetic-activated cell sorting, demonstrating vastly superior performance in selectivity, efficiency, and sorting speed. RESEARCH ARTiCLE
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荧光激活片上细胞培养分选(O3CS):智能培养皿
芯片上细胞分选是一种很有前途的干细胞培养分选技术。芯片上的细胞分选可以最大限度地减少实验室人员参与细胞处理,大大降低细胞培养污染的风险。我们开发了一种荧光激活的片上细胞培养分选(O3CS)系统,该系统结合了生物相容性半导体光寻址微阵列(芯片)和用于芯片寻址和细胞培养观察的光学装置。光学装置具有荧光和反射光显微镜功能,用于可视化和控制细胞群。基于光诱导电穿孔的高效分选器对“不需要的”细胞进行高分辨率检测是NeuroSyntek StemOptimizer 6+中实现的O3CS的核心。我们证明了该系统能够通过诱导不可逆的单细胞电穿孔来进行细胞培养荧光激活分选,并通过荧光显微镜和流式细胞术验证了O3CS分选的有效性,并将其与磁激活细胞分选进行了比较,结果表明该系统在选择性、效率和分选速度方面具有极大的优势。研究文章
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