Study of the manufacturing process and functionalization in order to achieve a vibrating micro device for specific detection in Biology
This study is a preliminary study to use a fonctionnalized microsensor based on electromechanical detection of biological species present at trace levels. Two aspects were involved in this work: capture high-sensitivity but also the capture of ultra-specific biological species as markers for Alzheimer's disease. It was conducted as part of collaboration between the department MINASYS (Micro and Nanobio and Microsystems) of the Institute of Fundamental Electronics (IEF) and the Laboratory Proteins and Nanotechnologies in Separative Sciences, at the Faculty of Pharmacy of Chatenay-Malabry.The sensor includes a micro-resonator, placed in vaccum, incorporating silicon buried channels for the circulation of a biological fluid and not outside as they are more commonly, and in order to minimize the damping of the resonator. As regards the biological aspects, we primily focus on the detection of a biomarker of Alzheimer's disease, amyloidal peptide Aβ1-42. Until now, screening requires the use of a puncture and cerebrospinal fluid after onset of symptoms, therefore a sufficiently high concentration of biomarkers is required. Initially, we developed a method of early detection of Aβ1-42 peptide coupling a microfluidic system and fluorescence microscopy. Therefore, the micro-resonator that are present here allow to detect these biomarkers that are present at very low concentrations in other biological fluids in the first stage of the disease, for example, in the blood or urine. Thus, in a first step, we have developed a method of early detection of Aβ1-42 peptide coupling channel microfluidic and fluorescence microscopy. The surface of the channels in silicon must be functionalized to allow the grafting of specific antigens. For this, we developed a technique based on the specific antibody-antigen recognition, the latter requires a preliminary step of chemical functionalization of surfaces.This manuscript consists of four main parts. Initially, we present a literature review of the state of the art on the principle of operation of various types commonly used bio-sensors and the performance obtained. A second chapter describes the functionalization of silicon area and more specifically of the silanization reaction in liquid phase performed on flat surfaces and in fluidic channels. In the following chapter, we discuss the field of specific recognition of biological entities and detail the steps of grafting performed on protein surfaces and design immunoassay sandwich in fluidic channels. The last chapter of the manuscript brings together various preliminary results for the development of micro-sensor type resonant beam.