微流控芯片技术用于精子与上皮细胞分离的研究

发布时间：2018-02-28 18:57

本文关键词： 微流控芯片精子上皮细胞段串联重复序列分离　出处：《中国细胞生物学学报》2013年10期 　论文类型：期刊论文

【摘要】：为研究建立使用微流控芯片技术分离精子与阴道上皮细胞的方法,选用制作工艺简单的玻璃-PDMS芯片,对混合样本进行分离。加样前分别在进、出口池中加入7μL和10μL缓冲液,然后在进样口加入2μL混合样本。至少静置8min后,从出口池中取出3μL形成重力驱动的微流体后开始分离,每隔5 min在进样口补加1μL缓冲液。达到理想分离效果时,用移液器从出口池取出分离出的精子,核酸酶去除游离DNA,经过提取、扩增和电泳分离/检测等步骤得到精子的分型结果。结果显示,使用基于重力驱动微流体原理的微流控芯片可在30 min内分离出精子,不会有上皮细胞进入分离通道;通过核酸酶对分离出的精子液的去游离DNA处理,可得到单一、完整的精子分型。与传统的差异裂解法相比,这种方法在很大程度上节省了检验时间,在性侵案件中具有一定的法医物证分析价值。
[Abstract]:In order to establish a method of separating spermatozoa and vaginal epithelial cells by microfluidic chip, a simple glass PDMS chip was used to separate mixed samples. 7 渭 L and 10 渭 L buffers were added to the inlet and outlet cisterns before adding the samples. Then 2 渭 L mixed sample was added at the injection port. After at least 8 minutes of statics, 3 渭 L gravity driven microfluid was removed from the exit tank and separated. 1 渭 L buffer solution was added at the injection port every 5 min. The ideal separation effect was achieved. The spermatozoa was extracted from the outlet pool with a liquid remover, and the free DNA was removed by nuclease. The results of sperm typing were obtained by extraction, amplification and electrophoresis separation / detection. Using microfluidic chip based on the principle of gravity driven microfluid, sperm can be separated within 30 min without any epithelial cells entering the separation channel. Compared with the traditional differential lysis method, this method saves the testing time to a great extent, and has certain value of forensic physical evidence analysis in sexual assault cases.
【作者单位】：公安部物证鉴定中心;北京理工大学生命学院;清华大学生物医学工程系;中国人民公安大学刑事科学技术学院;太原铁路公安局临汾公安处;
【基金】：公安部重点实验室项目(批准号:2011GABJC028)资助的课题~~
【分类号】：D919.4;TN47

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