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Phénotypage innovant de la coagulation in vitro par indicateur fluorescent : aspects photophysiques théoriques et innovation instrumentale avancée

Abstract : In situ investigation of thrombin activity generation consists in recording the hydrolysis of a thrombin substrate (either fluorigenic or chromogenic): it is named thrombography. That investigation allows an integrative in vitro phenotyping of the coagulation system, in presence of platelets and fibrin as far as the fluorigenic method is concerned, and its applications are hypo-, as well as hyper- coagulant states, even if they are the result of a complex coagulopathy disturbing the pro - and the anti -coagulant balance. The present work is the technological contribution of the INSERM unit U734, in collaboration with the Diagnostica Stago company. By means of a theoretical approach that describes the photophysical phenomena occurring during thrombography, a model (mathematical relation) based upon physical laws was obtained. It independently validated the calibration process of the CAT (Calibrated Automated Thrombography), which is currently the most relevant and used system for thrombography. The model can also be an alternative process for calibration, obtained by measuring the fluorescence at various but constant fluorophore concentrations, for the rare cases when the CAT calibration process is impaired (e.g. presence in the investigated sample of a direct thrombin inhibitor of low molecular weight). By means of an unitary disposable reagent container and innovative technological solutions in a multi-channel measurement bench, different from what is commercially available, an independent reproduction of the CAT results was obtained. By that combination, independent inputs / outputs in the cycle of measures for every used reagent container were made possible. Gauging of the independent channels was achieved by the alternative calibration. The experiment rate can be eventually increased compared to the available devices for thrombography and the human factor has to be decreased as much as possible by further automation
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Nicolas Lelievre. Phénotypage innovant de la coagulation in vitro par indicateur fluorescent : aspects photophysiques théoriques et innovation instrumentale avancée. Biologie cellulaire. Université Henri Poincaré - Nancy 1, 2008. Français. ⟨NNT : 2008NAN10042⟩. ⟨tel-01746309⟩

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