A microfluidic dual capillary probe to collect messenger RNA from adherent cells and spheroids
A microfluidic dual capillary probe to collect messenger RNA from adherent cells and spheroids
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DOI：
10.1016/j.ab.2008.10.039
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发表时间：
2009-02-01
影响因子：
2.9
通讯作者：
Matsue, Tomokazu
中科院分区：
文献类型：
--
作者：
Shiku, Hitoshi;Yamakawa, Takeshi;Matsue, Tomokazu
尽管最近在许多集成微流控装置中取得了进展，但从单细胞中收集生物分析物仍然是一项具有挑战性的技术。用θ形玻璃毛细管制备了微流控双毛细管探针，用于分析贴壁细胞和球状体的信使RNA（mRNA）。从注射孔引入细胞裂解缓冲液溶液，并收集来自抽吸孔的细胞裂解溶液用于基于逆转录实时PCR的进一步mRNA分析。通过使用微流体双毛细管探针，可以将细胞裂解缓冲液引入任何靶细胞处，并且不会溢出靶区域，因为在优化的注射/抽吸流速下在探针附近局部形成层流。该方法实现了在单细胞水平上对mRNA的敏感性，并根据相关基因表达谱对细胞类型进行鉴定。(C)2008年爱思唯尔公司All rights reserved.
Collection of bioanalytes from single cells is still a challenging technology despite the recent progress in many integrated microfluidic devices. A microfluidic dual capillary probe was prepared from a theta (theta)-shaped glass capillary to analyze messenger RNA (mRNA) from adherent cells and spheroids. The cell lysis buffer Solution was introduced from the injection aperture, and the cell-lysed solution from the aspiration aperture was collected for further mRNA analysis based on reverse transcription real-time PCR. The cell lysis buffer can be introduced at any targeted cells and never spilled out of the targeted area by using the microfluidic dual capillary probe because laminar flow was locally formed near the probe under the optimized injection/aspiration flow rates. This method realizes the sensitivity of mRNA at the single cell level and the identification of the cell types on the basis of the relative gene expression profiles. (C) 2008 Elsevier Inc. All rights reserved.