| Detailed view | pastel-00589602, version 1 |
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| Defence date | 2011-03-15 |
| Chimie, physico-chimie et génie chimique | |
| Library | Ecole Polytechnique |
| Keywords | Nanopores – Nanopore – Translocation – Bruit – Bruit rose |
| English title | Ions and Objects Transport in Nanopores |
| English abstract | Several ways exist to handle and detect single molecules. The oldest one is to measure the ionic current going through a single nanometric pore. This thesis uses this technique. The discussion is built in two main parts. Firstly, the use of a single track-etched nanopore is aimed to reproduce experiments already made in the litterature with other kinds of pores. Some phenomena, like rectification for example, were observed, whereas others, like translocation, were not. Secondly, the investigation of electric conductance noise proves the existence of a phenomenon which has not been pointed out before in the litterature yet. Indeed, this noise can be ascribed neither to fluctuations of the pore geometry nor to the pore wall charges, but rather to a cooperative effect on ions motion in confined geometry. |
| English keyword | Nanopores – Nanopore – Translocation – Noise – Pink Noise |
| Table des matières Introduction 1 1 Historique et état de l'art 3 1.1 Historique de la détection de molécule unique . . . . . . . . . 4 1.1.1 Pourquoi étudier une molécule unique ? . . . . . . . . . 4 1.1.2 Observation de particules sub-millimétriques . . . . . . 6 1.1.3 De particule unique à molécule unique . . . . . . . . . 8 1.2 Le nanopore naturel -hémolysine . . . . . . . . . . . . . . . . 11 1.2.1 Présentation de l' -hémolysine . . . . . . . . . . . . . 11 1.2.2 Translocation de macromolécules . . . . . . . . . . . . 13 La translocation en biologie . . . . . . . . . . . . . . . 13 Translocation de macromolécules non biologiques dans l' -hémolysine . . . . . . . . . . . . . . . . . 14 1.2.3 Translocation de polymères biologiques . . . . . . . . . 15 1.2.4 Avantages et limitations des nanopores naturels . . . . 17 Avantages . . . . . . . . . . . . . . . . . . . . . . . . . 17 Limitations . . . . . . . . . . . . . . . . . . . . . . . . 18 1.3 Les nanopores sur support silicium . . . . . . . . . . . . . . . 21 1.3.1 Développement . . . . . . . . . . . . . . . . . . . . . . 21 1.3.2 Translocation dans des pores sur support silicium . . . 24 Structure secondaire de l'ADN . . . . . . . . . . . . . . 24 Sens de variation de la conductance pendant la translocation . . . . . . . . . . . . . . . . . . . . . 27 Réduction de la vitesse de translocation . . . . . . . . 28 1.4 Les nanopores à trace attaquée . . . . . . . . . . . . . . . . . 31 1.4.1 Présentation . . . . . . . . . . . . . . . . . . . . . . . . 31 Taille et forme des pores . . . . . . . . . . . . . . . . . 32 Un pore unique . . . . . . . . . . . . . . . . . . . . . . 33 1.4.2 Translocation de macromolécules . . . . . . . . . . . . 34 ADN . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Protéines . . . . . . . . . . . . . . . . . . . . . . . . . 36 161 162 TABLE DES MATIÈRES 2 Préparation et caractérisation 39 2.1 Méthode de l'attaque de traces . . . . . . . . . . . . . . . . . 40 2.1.1 Nature du polymère . . . . . . . . . . . . . . . . . . . 40 Polycarbonate . . . . . . . . . . . . . . . . . . . . . . . 40 Polyimide . . . . . . . . . . . . . . . . . . . . . . . . . 41 2.1.2 Irradiation aux ions lourds . . . . . . . . . . . . . . . . 43 Irradiation avec un ion unique . . . . . . . . . . . . . . 48 2.1.3 Attaque chimique . . . . . . . . . . . . . . . . . . . . . 49 Modèle à deux vitesses d'attaque . . . . . . . . . . . . 49 Suivi par conductimétrie . . . . . . . . . . . . . . . . . 49 Attaque symétrique de trace unique dans le polycarbonate . . . . . . . . . . . . . . . . . . . . . . 51 Attaque asymétrique de trace unique dans le polyimide 55 2.2 Caractérisation de la géométrie . . . . . . . . . . . . . . . . . 57 2.2.1 Contexte . . . . . . . . . . . . . . . . . . . . . . . . . . 57 2.2.2 Microscopie électronique . . . . . . . . . . . . . . . . . 57 Détermination de v? dans le polycarbonate . . . . . . . 58 Grand rayon des pores coniques dans le polyimide . . . 61 2.2.3 Conductimétrie . . . . . . . . . . . . . . . . . . . . . . 62 Dispositif expérimental . . . . . . . . . . . . . . . . . . 62 Détermination du petit rayon . . . . . . . . . . . . . . 64 Limites de la conductimétrie . . . . . . . . . . . . . . . 67 2.3 Caractérisation de la paroi du pore . . . . . . . . . . . . . . . 68 2.3.1 Phénomène de recti cation . . . . . . . . . . . . . . . . 68 Courbe intensité-tension . . . . . . . . . . . . . . . . . 68 Non-linéarité des courbes I-V . . . . . . . . . . . . . . 69 2.3.2 Comportement en milieu acide . . . . . . . . . . . . . . 69 2.3.3 Potentiel d'écoulement . . . . . . . . . . . . . . . . . . 72 Double couche électrique et potentiel . . . . . . . . . 72 Calcul du potentiel d'écoulement . . . . . . . . . . . . 74 Expériences . . . . . . . . . . . . . . . . . . . . . . . . 77 Variations attendues du potentiel avec la concentration 79 Variations du potentiel avec la taille du pore . . . . . 81 3 Transport des ions 83 3.1 Recti cation . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 3.1.1 Charge surfacique sur la paroi interne . . . . . . . . . . 84 3.1.2 Longueur de Debye . . . . . . . . . . . . . . . . . . . . 86 3.1.3 Modélisation . . . . . . . . . . . . . . . . . . . . . . . . 87 3.1.4 La recti cation dans le cadre de cette thèse . . . . . . 90 3.2 E ets de con nement . . . . . . . . . . . . . . . . . . . . . . . 92 TABLE DES MATIÈRES 163 3.2.1 Exemple de l' -hémolysine . . . . . . . . . . . . . . . . 92 3.2.2 Le con nement dans un pore à trace attaquée . . . . . 94 Notations . . . . . . . . . . . . . . . . . . . . . . . . . 94 Variations avec la concentration en électrolyte . . . . . 95 3.3 Bruit de conduction électrique . . . . . . . . . . . . . . . . . . 99 3.3.1 Densité spectrale de puissance . . . . . . . . . . . . . . 99 Le bruit rose . . . . . . . . . . . . . . . . . . . . . . . 99 3.3.2 Variations du bruit selon l'intensité . . . . . . . . . . . 102 3.3.3 Origines du bruit rose . . . . . . . . . . . . . . . . . . 103 3.3.4 Liquides ioniques . . . . . . . . . . . . . . . . . . . . . 105 3.3.5 Les liquides ioniques et le bruit rose . . . . . . . . . . . 107 3.3.6 Bruit rose et con nement . . . . . . . . . . . . . . . . . 109 Origines du bruit rose : pore ou solution ? . . . . . . . 109 4 Translocation et transport d'objets 115 4.1 Translocation de la luciférase . . . . . . . . . . . . . . . . . . 116 4.1.1 Présentation de l'aérolysine . . . . . . . . . . . . . . . 116 4.1.2 La luciférase, une protéine adaptée . . . . . . . . . . . 117 Présentation . . . . . . . . . . . . . . . . . . . . . . . . 117 Mesure de la concentration . . . . . . . . . . . . . . . . 118 Dénaturation de la luciférase . . . . . . . . . . . . . . . 120 4.1.3 Aérolysine et luciférase dénaturée . . . . . . . . . . . . 122 Dispositif et protocole expérimentaux . . . . . . . . . . 122 Observation de translocations par conductimétrie . . . 123 4.2 Tentatives de transport . . . . . . . . . . . . . . . . . . . . . . 126 4.2.1 Protéines . . . . . . . . . . . . . . . . . . . . . . . . . 126 Luciférase . . . . . . . . . . . . . . . . . . . . . . . . . 126 BSA (Bovine Serum Albumin) [Albumine de sérum bovin] . . . . . . . . . . . . . . . . . . . . . . . 127 Fibronectine . . . . . . . . . . . . . . . . . . . . . . . . 127 4.2.2 ADN . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 4.2.3 Nanoparticules . . . . . . . . . . . . . . . . . . . . . . 128 4.2.4 Polymères et polyélectrolytes . . . . . . . . . . . . . . 129 4.2.5 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . 129 Tension appliquée . . . . . . . . . . . . . . . . . . . . . 130 Concentration en objets . . . . . . . . . . . . . . . . . 130 Taille des objets et du pore . . . . . . . . . . . . . . . 130 Interactions avec la paroi . . . . . . . . . . . . . . . . . 131 Conclusion générale 133 164 TABLE DES MATIÈRES Bibliographie 135 Index 149 Table des gures et des tableaux 151 Table des sigles et abbréviations 159 |
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