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Fluorescence Microscopy and Microfluidics for Cellular and Enzymatic Analysis

  • Author(s): Walsh, Matthew Thomas
  • et al.
Abstract

Breakthroughs in our understanding of biological systems are driven by technological advancements that enable the study of cellular and molecular processes at unprecedented levels. Recent efforts have provided a thrust in the development of microfluidic devices that facilitate the process of genomic or transcriptomic library construction from single cells. The integration of molecular detection modalities within microfluidic devices will enable the direct analysis of cellular content through surface capture and probing of biomolecules. Similar techniques can be applied to monitoring the enzymatic reaction processes in a rapid and high-throughput manner. We have demonstrated the application of fluorescence microscopy and microfluidics to perform the automatic hydrodynamic capture of single mammalian cells followed by the immobilization and digital counting of polyadenylated mRNA molecules released from the individual cells. Fluorescence imaging was also utilized to monitor the incorporation kinetics of nucleotides by DNA polymerase within a microfluidic device using multiple dye-labeling strategies. The combination of advanced microfluidic devices and sensitive fluorescence detection techniques provides a powerful tool set for enabling sophisticated cell and molecular biology experimentation

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