Multi-color fluorescent DNA analysis in an integrated optofluidic lab-on-a-chip


Dongre, C. and Weerd, J. van and Weeghel, R. van and Martinez Vazquez, R. and Osellame, R. and Cerullo, G. and Besselink, G.A.J. and Vlekkert, H.H. van den and Hoekstra, H.J.W.M. and Pollnau, M. (2010) Multi-color fluorescent DNA analysis in an integrated optofluidic lab-on-a-chip. In: Sense of Contact 12, April 8, 2010, Zeist, The Netherlands.

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Abstract:Sorting and sizing of DNA molecules within the human genome project has enabled the genetic mapping of various illnesses. By employing tiny lab-on-a-chip devices for such DNA analysis, integrated DNA sequencing and genetic diagnostics have become feasible. However, such diagnostic chips typically lack integrated sensing capability. We address this issue by combining microfluidic capillary electrophoresis with laser-induced fluorescence detection resulting in optofluidic integration towards an on-chip bio-analysis tool [1,2]. We achieve a spatial separation resolution of 12 μm, which can enable a 20-fold enhancement in electropherogram peak resolution, leading to plate numbers exceeding one million. We demonstrate a high sizing/calibration accuracy of 99% [3], and ultrasensitive fluorescence detection (limit of detection = 65 femtomolar, corresponding to merely 2-3 molecules in the excitation/detection volume) of diagnostically relevant double-stranded DNA molecules by integrated-waveguide laser excitation. Subsequently, we introduce a principle of parallel optical processing to this optofluidic lab-on-a-chip. Different sets of exclusively color-labeled DNA fragments – otherwise rendered indistinguishable by their spatio-temporal coincidence – are traced back to their origin by modulation-frequency-encoded multi-wavelength laser excitation, fluorescence detection with a color-blind photomultiplier, and Fourier-analysis decoding. As a proof of principle, fragments from independent human genomic segments, associated with genetic predispositions to breast cancer and anemia, are extracted by multiplex ligation-dependent probe amplification, and simultaneously analyzed. Such multiple yet unambiguous optical identification of biomolecules opens new horizons for “enlightened” lab-on-a-chip devices.
Item Type:Conference or Workshop Item
Electrical Engineering, Mathematics and Computer Science (EEMCS)
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