On-chip cell lysis by local hydroxide generation
On-chip cell lysis by local hydroxide generation
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DOI：
10.1039/b413139h
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发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Lee, LP
中科院分区：
文献类型：
--
作者：
Di Carlo, D;Ionescu-Zanetti, C;Lee, LP
我们提出了一种基于局部氢氧化物电生成的芯片上细胞裂解的新方法。氢氧根离子穿过细胞膜，导致细胞裂解。裂解发生后，芯片上也会产生氢离子，与过量的氢氧化物反应，产生中性 pH 裂解液，从而无需清洗步骤。该方法可有效裂解三种不同的细胞类型：红细胞、HeLa（人类肿瘤系）和中国仓鼠卵巢 (CHO) 细胞系。通过监测膜不透性染料从细胞质中的逃逸来证明 HeLa 和 CHO 细胞释放细胞质分子。在阴极附近，氢氧化物浓度通过有限元模拟进行预测，并显示适合距产生阴极不同距离处的裂解速率。对于流通实验，制造并测试了将机械过滤器与氢氧化物生成集成的第二个装置。过滤器的目的是捕获整个细胞并只允许裂解物通过。对裂解过滤器处氢氧化物浓度的流速依赖性进行了建模，并且通过实验确定裂解效率与流速从 15 到 30 mul min(-1) 的氢氧化物浓度成正比。
We present a novel method for on-chip cell lysis based on local hydroxide electro-generation. Hydroxide ions porate the cell membrane, leading to cell lysis. After lysis occurs, hydrogen ions, also generated on chip, react with excess hydroxide, creating a neutral pH lysate and eliminating the need for a wash step. Three different cell types are shown to be effectively lysed by this method: red blood cells, HeLa ( human tumor line) and Chinese Hamster Ovary (CHO) cell lines. The release of cytoplasmic molecules from HeLa and CHO cells is demonstrated by monitoring the escape of a membrane impermeant dye from the cytoplasm. In the vicinity of the cathode, the hydroxide concentration is predicted by finite element simulations and shown to fit the lysis rates at different distances from the generating cathode. For flow-through experiments, a second device integrating a mechanical filter with hydroxide generation is fabricated and tested. The purpose of the filter is to trap whole cells and only allow lysate to pass through. The flow rate dependence of hydroxide concentration at the lysis filter is modeled and lysis efficiency is experimentally determined to be proportional to the hydroxide concentration for flow rates from 15 to 30 mul min(-1).