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首页> 外文期刊>RSC Advances >A 3D mixing-based portable magnetic device for fully automatic immunofluorescence staining of γ-H2AX in UVC-irradiated CD4+ cells
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A 3D mixing-based portable magnetic device for fully automatic immunofluorescence staining of γ-H2AX in UVC-irradiated CD4+ cells

机译：基于3D混合的便携式磁性装置，用于UVC辐照的CD4 +细胞中γ-H2AX的全自动免疫荧光染色

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Immunofluorescence (IF) is a common method used in cell biology. The conventional protocol for IF staining is time and labor-intensive, operator dependent and reagent-consuming. Magnetic Bead (MB)-based microdevices are frequently utilized in cellular assays, but integration of simple and efficient mixing with downstream multi-step manipulation of MBs for automatic IF staining is still challenging. We herein present a portable, inexpensive and integratable device for MB-based automatic IF staining. First, a front-end cell capture step is performed using a 3D-mixing module, which is built upon a novel mechanism named ec-2MagRotors and generates periodically changing 3D magnetic fields. A 5-fold enhancement of cell capture efficiency was attained even with a low bead-to-cell concentration ratio (5?:?1), when conducting magnetic 3D mixing. Second, a 1D-moving module is employed downstream to automatically manipulate MB–cell complexes for IF staining. Further, a simplified protocol for staining of γ-H2AX, a biomarker widely used in evaluation of cell radiation damage, is presented for proof-of-principle study of the magnetic device. Using UVC-irradiated CD4 ~(+) cells as samples, our device achieved fully automatic γ-H2AX staining within 40 minutes at room temperature and showed a linear dose–response relationship. The developed portable magnetic device is automatic, efficient, cost-effective and simple-to-use, holding great potential for applications in different IF assays.
机译：免疫荧光（IF）是在细胞生物学中使用的常用方法。对于IF染色的常规协议是时间和劳动力密集型的，依赖于操作者和试剂消耗。磁珠（MB）为基础的微型器件经常用于在细胞试验，但简单和有效的混合与自动MB的下游多步操作的集成IF染色仍然具有挑战性。本文我们提出了基于MB-自动IF染色的便携式的，廉价的，可集成的设备。首先，使用3D-混合模块，其被命名为在EC-2MagRotors一种新的机制建立并产生周期性变化的三维磁场，进行前端细胞捕获步骤。的细胞捕获效率的5倍的增强物获得即使在低珠 - 细胞浓度比（5：？1），进行磁3D混合时。第二，一维移动的模块采用下游自动操纵MB - 细胞复合物为IF染色。此外，对于γ-H2AX，广泛用于细胞辐射损伤评价的生物标志物的染色的简化协议，提出了一种用于磁性器件的验证的原理研究。使用UVC照射CD4〜（+）细胞作为样品，我们的装置来实现全自动γ-H2AX在室温下40分钟内染色和呈线性剂量 - 反应关系。的开发的便携式磁性装置是自动的，高效的，成本有效和简单易用的，为应用程序在不同的IF测定保持很大的潜力。

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