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

Hitoshi Shiku, Takeshi Yamakawa, Yuji Nashimoto, Yasufumi Takahashi, Yu suke Torisawa, Tomoyuki Yasukawa, Takahiro Ito-Sasaki, Masaki Yokoo, Hiroyuki Abe, Hideki Kambara, Tomokazu Matsue

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)138-142
Number of pages5
JournalAnalytical Biochemistry
Volume385
Issue number1
DOIs
Publication statusPublished - 2009 Feb 1

Keywords

  • Microfluidics
  • RT-PCR
  • Real-time PCR
  • Single cell analysis

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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