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http://hdl.handle.net/10397/88188
Title: | A 3D mixing-based portable magnetic device for fully automatic immunofluorescence staining of gamma-H2AX in UVC-irradiated CD4(+)cells | Authors: | Zhong, RT Hou, LS Zhao, YB Wang, TL Wang, SH Wang, MY Xu, D Sun, YQ |
Issue Date: | 2020 | Source: | RSC advances, 2020, v. 10, no. 49, p. 29311-29319 | Abstract: | 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 gamma-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 gamma-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. | Publisher: | Royal Society of Chemistry | Journal: | RSC advances | EISSN: | 2046-2069 | DOI: | 10.1039/d0ra03925j | Rights: | This journal is © The Royal Society of Chemistry 2020 This article is licensed under a Creative Commons Attibution-NonCommercial 3.0 Unported Licence (https://Creative Commons.org/licenses/by-nc/3.0/) The following publication Zhong, R. T., Hou, L. S., Zhao, Y. B., Wang, T. L., Wang, S. H., Wang, M. Y., . . . Sun, Y. Q. (2020). A 3D mixing-based portable magnetic device for fully automatic immunofluorescence staining of gamma-H2AX in UVC-irradiated CD4(+)cells. RSC advances, 10(49), 29311-29319 is available at https://dx.doi.org/10.1039/d0ra03925j |
Appears in Collections: | Journal/Magazine Article |
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