Analysis of gene expression at the single-cell level using microdroplet-based microfluidic technology
Analysis of gene expression at the single-cell level using microdroplet-based microfluidic technology
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DOI：
10.1063/1.3596394
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发表时间：
2011-06-01
期刊：
影响因子：
3.2
通讯作者：
Tabeling, Patrick
中科院分区：
文献类型：
--
作者：
Mary, Pascaline;Dauphinot, Luce;Tabeling, Patrick
在目前的工作中，我们测量了总 RNA 和封装在液滴中的细胞中特定基因的信使 RNA 表达。推出的微流控芯片包括以下功能：RNA/细胞封装、裂解、逆转录和实时聚合酶链式反应。我们已经证明，可以在不到 2 小时内对同时封装在 2 nl 液滴中的 100 个纯化 RNA 样本进行单链和双链基因表达测量。对包含一到三个细胞的 100 个样品的分析表明，平均值与标准技术具有极好的一致性。 E-钙粘蛋白表达的细胞间分布表明转录本数量的波动约为 80%，这与文献中的一般发现高度一致。还引入了数学模型来加强对我们结果的解释。目前的工作为生物和生物医学研究中系统获取此类信息铺平了道路。 (C) 2011 年美国物理研究所。 [doi:10.1063/1.3596394]
In the present work, we have measured the messenger RNA expression of specific genes both from total RNA and cells encapsulated in droplets. The microfluidic chip introduced includes the following functionalities: RNA/cell encapsulation, lysis, reverse transcription and real-time polymerase chain reaction. We have shown that simplex and duplex gene expression measurements can be carried out over a population of 100 purified RNA samples encapsulated simultaneously in 2 nl droplets in less than 2 h. An analysis of 100 samples containing one to three cells has shown excellent consistency with standard techniques regarding average values. The cell-to-cell distributions of the E-cadherin expression suggest fluctuations on the order of 80% in the number of transcripts, which is highly consistent with the general findings from the literature. A mathematical model has also been introduced to strengthen the interpretation of our results. The present work paves the way for the systematic acquisition of such information in biological and biomedical studies. (C) 2011 American Institute of Physics. [doi:10.1063/1.3596394]