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

C. Dongre, J. van Weerd, R. van Weeghel, R. Martinez-Vazquez, R. Osellame, G. Cerullo, G.A.J. Besselink, H.H. van den Vlekkert, Hugo Hoekstra, Markus Pollnau

    Research output: Chapter in Book/Report/Conference proceedingConference contributionAcademic

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    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.
    Original languageEnglish
    Title of host publicationProceedings of Sense of Contact 12
    Place of PublicationZeist
    PublisherFHI - Federatie Van Technologiebranches
    Number of pages1
    ISBN (Print)978-90-74702-56-0
    Publication statusPublished - Apr 2010
    Event12th Sensor Technology Conference Sense of Contact 2010 - Conferentiecentrum Woudschoten, Zeist, Netherlands
    Duration: 8 Apr 20108 Apr 2010
    Conference number: 12


    Workshop12th Sensor Technology Conference Sense of Contact 2010




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