Objetivo Currently, the two prominent schemes for single-molecule fluorescence detection (SMFD), confocal microscopy and camera-based total-internal-reflection or wide-field microscopy, are ultimately limited in their ability to combine the detection of many molecules with obtaining data at sufficiently high time resolution, necessary for resolving fast dynamics with single-molecule fluorescence resonance energy transfer (smFRET). In particular, monitoring enzymatic reactions using smFRET is extremely challenging and remains to a large extent unexplored.Here, I propose a novel nanofluidic device to overcome these limitations by using nanochannels, which provide a well-defined flow path for afluorescent species through the excitation/detection focus of a conventional wide-field microscope. Using an array of nanochannels offers several advantages: First, the geometrical confinement enables long observation times of non-immobilized molecules. Second, the residence time of molecules in the channels is easily controlled by the flow velocity set by the syringe pump. Third, faster flow rates together with using a CCD camera in ‘streaking mode’ enable a sub-millisecondtime resolution. Fourth, a high-throughput detection is achieved by using a parallel array of channels. Fifth, enzymatic reactions can be directly triggered by mixing necessary components on-the-fly using an additional inlet.The described device will pave the way for high-throughput single-molecule detection, which will greatly expand the possibilities for researchers to apply single-molecule methods in the area of Life Sciences. Ámbito científico engineering and technologyelectrical engineering, electronic engineering, information engineeringelectronic engineeringsensorsoptical sensorsnatural sciencesphysical sciencesopticsmicroscopyconfocal microscopy Programa(s) FP7-PEOPLE - Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Tema(s) FP7-PEOPLE-2013-CIG - Marie-Curie Action: "Career Integration Grants" Convocatoria de propuestas FP7-PEOPLE-2013-CIG Consulte otros proyectos de esta convocatoria Régimen de financiación MC-CIG - Support for training and career development of researcher (CIG) Coordinador WAGENINGEN UNIVERSITY Aportación de la UE € 100 000,00 Dirección DROEVENDAALSESTEEG 4 6708 PB Wageningen Países Bajos Ver en el mapa Región Oost-Nederland Gelderland Veluwe Tipo de actividad Higher or Secondary Education Establishments Contacto administrativo Tom Bessems (Mr.) Enlaces Contactar con la organización Opens in new window Sitio web Opens in new window Coste total Sin datos
