Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip
| Autor: | Jasper van Weerd, Hans H. van den Vlekkert, C. Dongre, R.M. Vazquez, Roberto Osellame, Giulio Cerullo, Hugo Hoekstra, Markus Pollnau, G.A.J. Besselink, Rob van Weeghel |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Electrophoresis
Photomultiplier Biomedical Engineering Bioengineering Nanotechnology Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip Biochemistry Sensitivity and Specificity DNA sequencing law.invention chemistry.chemical_compound Capillary electrophoresis law Lab-On-A-Chip Devices Humans Multiplex Oligonucleotide Array Sequence Analysis Physics Fourier Analysis business.industry General Chemistry DNA Laser Fluorescence Spectrometry Fluorescence chemistry IOMS-SNS: SENSORS Optoelectronics business Frequency modulation |
| Zdroj: | International Laser Physics Workshop 2011 Lab on a chip 11 (2011): 679–683. doi:10.1039/c0lc00449a info:cnr-pdr/source/autori:C. Dongre, J. van Weerd, G.A.J. Besselink, R. Martinez Vazquez, R. Osellame, G. Cerullo, R. van Weeghel, H.H. van den Vlekkert, H.J.W.M. Hoekstra, M. Pollnau/titolo:Modulation-frequency encoded multi-color fluorescent DNA analysis in an optofluidic chip/doi:10.1039%2Fc0lc00449a/rivista:Lab on a chip (Print)/anno:2011/pagina_da:679/pagina_a:683/intervallo_pagine:679–683/volume:11 Lab on a chip, 11(4), 679-683. Royal Society of Chemistry |
| ISSN: | 1473-0197 |
| DOI: | 10.1039/c0lc00449a |
| Popis: | By capillary electrophoresis (CE) in miniaturized lab-on-a-chip devices, integrated DNA sequencing and genetic diagnostics have become feasible. We introduce a principle of parallel optical processing to significantly enhance analysis capabilities. In a commercial microfluidic chip, a plug of DNA molecules was injected and the DNA molecules were CE-separated with a high relative sizing accuracy of >99%. Through an optical waveguide inscribed by femtosecond-laser writing a laser was launched perpendicularly into the microfluidic channel. A photomultiplier collected the fluorescence signals from a small detection window with a limit of detection of ~8 DNA molecules. In our approach, different sets of exclusively end-labeled DNA fragments are unambiguously identified by simultaneously launching several continuous-wave lasers, each modulated with a different frequency, detection of the frequency-encoded signals at different fluorescence wavelengths by a single ultrasensitive, albeit color-blind photomultiplier, and Fourier-domain frequency decoding. As a proof of principle, fragments from independent human genomic segments, associated with genetic predispositions to breast cancer and anemia, are simultaneously analyzed in a single flow experiment. This novel method of modulation-frequency-encoded fluorescence excitation opens new opportunities in genetic diagnostics. It enables the identification of end-labeled DNA samples of different genetic origin during their electrophoretic separation, opening perspectives for intrinsic size calibration, malign / healthy sample comparison, and exploitation of multiplex ligation-dependent probe amplification. |
| Databáze: | OpenAIRE |
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