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Arnaud Rival

Digital microfluidics for sample and gene expression analysis by qRT-PCR

Published on 26 November 2010
Thesis presented November 26, 2010

Digital microfluidics, in particular electrowetting on dielectric (EWOD), is an efficient method for dispensing and manipulating nanoliters droplets. These droplets represent reaction mediums with reduced volumes, specially adapted to complex biological issues, usually difficult to handle in larger volumes. We used​ EWOD technology to develop a lab-on-a-chip dedicated to gene expression studies on tiny biological samples.
Microsystem fabrication steps, such as processing of dielectric and hydrophobic layers, were optimized to operate at 95 °C. A specific test bench was also developed. All the necessary fluidic operations for qRT-PCR protocol were characterized, optimized and controlled. For instance, two mixing modes inside droplets were pointed out and studied in detail. Moreover, we demonstrated the ability of EWOD technology to handle magnetic beads on chip. Finally, this lab-on-a-chip was used to validate a protocol of transcriptome analysis starting from a few cells and even a single cell, integrating messenger RNAs extraction followed their analysis by qRT-PCR on a same chip.

Lab-on-a-chip, digital microfluidics, electrowetting, EWOD, sample preparation, gene expression, transcriptome, single cell.