ac5b03875_si_006.mp4 (1.28 MB)
Integration of Multiplexed Microfluidic Electrokinetic Concentrators with a Morpholino Microarray via Reversible Surface Bonding for Enhanced DNA Hybridization
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posted on 2016-02-25, 00:00 authored by Diogo Martins, Xi Wei, Rastislav Levicky, Yong-Ak SongWe describe a microfluidic concentration
device to accelerate the
surface hybridization reaction between DNA and morpholinos (MOs) for
enhanced detection. The microfluidic concentrator comprises a single
polydimethylsiloxane (PDMS) microchannel onto which an ion-selective
layer of conductive polymer poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate)
(PEDOT:PSS) was directly printed and then reversibly surface bonded
onto a morpholino microarray for hybridization. Using this electrokinetic
trapping concentrator, we could achieve a maximum concentration factor
of ∼800 for DNA and a limit of detection of 10 nM within 15
min. In terms of the detection speed, it enabled faster hybridization
by around 10-fold when compared to conventional diffusion-based hybridization.
A significant advantage of our approach is that the fabrication of
the microfluidic concentrator is completely decoupled from the microarray;
by eliminating the need to deposit an ion-selective layer on the microarray
surface prior to device integration, interfacing between both modules,
the PDMS chip for electrokinetic concentration and the substrate for
DNA sensing are easier and applicable to any microarray platform.
Furthermore, this fabrication strategy facilitates a multiplexing
of concentrators. We have demonstrated the proof-of-concept for multiplexing
by building a device with 5 parallel concentrators connected to a
single inlet/outlet and applying it to parallel concentration and
hybridization. Such device yielded similar concentration and hybridization
efficiency compared to that of a single-channel device without adding
any complexity to the fabrication and setup. These results demonstrate
that our concentrator concept can be applied to the development of
a highly multiplexed concentrator-enhanced microarray detection system
for either genetic analysis or other diagnostic assays.
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