Abstract
With a view to developing a rapid pathogen detection system utilizing isothermal nucleic acid amplification, the necessary micro-mixing step is innovatively implemented on a chip. Passive laminar flow mixing of two 6.5 μl batches differing in viscosity is performed within a microfluidic chamber. This is achieved with a novel chip space-saving phaseguide design which allows, for the first time, the complete integration of a passive mixing structure into a target chamber. Sequential filling of batches prior to mixing is demonstrated. Simulation predicts a reduction of diffusive mixing time from hours down to one minute. A simple and low-cost fabrication method is used which combines dry film resist technology and direct wafer bonding. Finally, an isothermal nucleic acid detection assay is successfully implemented where fluorescence results are measured directly from the chip after a one minute mixing sequence. In combination with our previous work, this opens up the way towards a fully integrated pathogen detection system in a lab-on-a-chip format.
Highlights
Lab-on-a-chip solutions have been recognised as an opportunity to bring accurate and sensitive diagnostic tests to the point-ofcare (POC).[1]
For a complete integration of the latest innovative assays, e.g. recombinase polymerase amplification2 (RPA),[2] unit operations such as micro-mixing need to be implemented on chip
In microfluidics low Reynolds numbers are common due to the small dimensions and the fluidics are dominated by laminar flow.[4]
Summary
Lab-on-a-chip solutions have been recognised as an opportunity to bring accurate and sensitive diagnostic tests to the point-ofcare (POC).[1]. A passive microfluidic batch mixing chip is presented which is based on the recently introduced phaseguide technology.[18] complete control over the filling and emptying of any type of microfluidic structures is given. This allows, for the first time, the direct integration of a mixing structure into a functional microfluidic chamber, such as the detection chamber of an assay. The applicability of this system is demonstrated with isothermal recombinase polymerase amplification[2] (RPA) on chip including its necessary mixing step. This detection assay, in combination with the ease of integration of phaseguided fluid handling, is promising with a view to a new generation of point of care diagnostic systems
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