A microfluidic dual capillary probe to collect messenger RNA from adherent cells and spheroids

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• 作者：Hitoshi Shiku ; Takeshi Yamakawa ; Yuji Nashimoto ; Yasufumi Takahashi ; Yu-suke Torisawa ; Tomoyuki Yasukawa ; Takahiro Ito-Sasaki ; Masaki Yokoo ; Hiroyuki Abe ; Hideki Kambara ; Tomokazu Matsue
• 关键词：RT-PCR ; Real-time PCR ; Microfluidics ; Single cell analysis
• 刊名：Analytical Biochemistry
• 出版年：2009
• 出版时间：1 February 2009
• 年：2009
• 卷：385
• 期：1
• 页码：138-142
• 全文大小：434 K

文摘

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 (θ)-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.