基于等电场平方梯度介电泳的高分辨率多细胞分选方法研究

Cell sorting has great research and application value in the area of biomedicine and life science, especially involving multi-cell sorting in complex samples, which is of significance to cell detection and analysis At present, most label-free cell sorting methods are limited in the number of the cell types and comprehensive performance. In this proposal, a sorting method based on constant electric field square gradient (CEFSG) dielectrophoresis is proposed to achieve precise sorting of various cells (such as four or more types of cells) by increasing the resolution of cell sorting. The main research contents include: The construction of CEFSG electric field based on photoconductive system, the dynamic model of various cells under CEFSG dielectrophoresis, the key influencing factors of the separation resolution of CEFSG dielectric method, and the precise sorting of multiple types of cell samples. In order to overcome the deficiency of the square gradient distribution and poor sorting effect of the traditional dielectrophoresis electric field, the optical conductivity system is used to construct the CEFSGelectric field, so that the separation force of the cells is only related to the characteristics of the cells themselves, thus significantly improving the sorting resolution and realizing the precise sorting of multiple types of cells. This method has characteristics of free-labeling, high resolution and accurate sorting of multiple cells, which provides strong support for cell detection, and is significant to the development of biomedical related fields and the cross-integration of medical disciplines.

细胞分选在生物医学和生命科学中具有重要的研究与应用价值，尤其是复杂样本中的多细胞分选对细胞检测与分析具有重要意义。目前多数免标记的细胞分选方法无论在分选种类，还是综合性能方面，均具一定局限性。本课题提出了一种基于等电场平方梯度（CEFSG）介电泳的分选方法，通过提高细胞分选分辨率来实现多种（如四种或以上）细胞的精准分选。主要研究内容包括：基于光电导体系CEFSG电场的构建；多种细胞在CEFSG介电力作用下的动力学模型；CEFSG介电法分选分辨率的关键影响因素；多细胞样本精准分选。为了克服传统介电泳电场平方梯度分布不均、分选效果差的不足，利用光电导体系构建了等电场平方梯度电场，使细胞受到的分选力仅与细胞本身特性有关，由此显著提高分选分辨率，实现多细胞精准分选。本方法具有免标记、分辨率高、多细胞精准分选等特点，可为细胞检测提供有力支撑，对生物医学相关领域的发展和医工学科交叉融合具有重要意义。

细胞是维持生物生命活动的基本单位，细胞、核酸、蛋白质等是生物医学领域最主要的研究对象，而细胞分选是细胞研究中重要的研究方法之一。本课题完成了一种基于等电场平方梯度（CEFSG）介电泳的多种细胞高分辨率无标记分选方法。本项目首先构建了电场数值物理模型与梯度下降法结合的快速、自动化求解CEFSG电场分布的方法；完成了光电导微流控芯片的加工，基于此芯片实现了CEFSG电场，并进行了验证。其次完成了不同细胞的介电泳模型的建立，研究了不同细胞、不同悬浮液、不同信号频率下细胞受到的介电泳力；建立了细胞在溶液中介电泳力作用下的细胞动力学模型，其为一个与介电泳力成正比的准静态过程；基于上述两个结论，得出细胞在一定的频率范围内，在介电泳力作用下可以实现分选；研究了信号幅值、微流道高度等因素对细胞分选分辨率的影响，给出了细胞分选建议的参数。最后利用CEFSG介电力细胞分选系统，完成了四种细胞的分选，提出了多频分选方法：在100kHz时，死细胞和活细胞的移动速度有显著性差别，因此可以实现活死细胞的分选；在300kHz时，Hela和MCF-7有显著性的速度差异，实现了两种活细胞的分选。综上所述，CEFSG介电泳提供了一种实现多种细胞和活死细胞的高分辨率无标记分选方法。基于此技术可以实现多种细胞分选相关生物医疗仪器的开发，对生物医学相关领域的发展和医工学科交叉融合具有重要意义。

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