Thermal microsensors for in vitro and in vivo monitoring of chemical and biological processes

Metabolism is a highly coordinated biochemical cellular activity indispensable for sustaining life, and is often related to mitochondrial functioning. Innovative technological approaches allowing the study of the thermal signature of chemical and metabolic processes are presented in this thesis. In particular, two solutions are proposed: (i) a nanocalorimetric platform enabling measurements of biochemical heat production in vitro, and (ii) a method for localized in vivo temperature measurements in mice using implantable miniaturized sensors. First, the design and development of the nanocalorimetric platform is presented. In particular, this platform is built around a commercial thermopile-based sensor chip, on which an open-well reservoir holding a sample with a volume of a few tens of microliter is directly positioned. Both components are embedded into an isothermal housing providing an excellent temperature stability (± 1 mK), which is a prerequisite for accurate heat power measurements. Furthermore, the platform is characterized by a fast thermalization time to the target temperature (less than 30 minutes) and a fast sensing response time (a few seconds). Numerical simulations, which allowed the optimization of the platform design, are also reported. Electrical calibration of the platform was carried out using resistive heaters positioned either on the sensor chip or in the sample reservoir. A heat power limit of detection of 70 nW was estimated. Then, two reactions for the validation of the nanocalorimetric platform were investigated: (i) the mixing of 1-propanol in water, and (ii) the oxidation of glucose catalyzed by glucose oxidase. The results showed good agreement with literature, confirming that our versatile platform may be applied to many thermochemical studies, including thermodynamic analysis and kinetic reaction analysis. Furthermore, localized in vivo temperature measurements in mice by means of implantable miniaturized sensors are presented. The aim was to monitor mouse metabolism during cold exposure, and to record possible temperature differences between the body temperature measured in the abdomen and the temperature of the brown adipose tissue (BAT) situated in the interscapular area. This approach is of biological interest as it may help unravelling the question whether biochemical activation of BAT is associated with local increase in metabolic heat production. Specifically, the preparation and calibration of miniaturized thermistor sensors are reported and the surgical procedure is also described. Temperature measurements, carried out on mice during cold exposure (6 °C) for a maximum duration of 6 hours, are presented and analyzed. Control measurements with a conventional probe confirmed the good performance of the implanted sensors. Moreover, analysis of the experimental results allowed the identification of two different mouse phenotypes, distinguishable in terms of their metabolic resistance to cold exposure. This difference was investigated from the thermal point of view by computational simulations. A simple physical model of the mouse body allowed to reproduce the global evolution of hypothermia and to explain qualitatively the temperature difference between abdomen and BAT locations. While with this approach, we have demonstrated the importance and feasibility of localized temperature measurements on mice, further optimization of this technique is foreseen to better identify local metabolism variations.

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