Patterning of tethered bilayer lipid membranes [Elektronische Ressource] / Mathieu Jung

Jung

Sujets

• Biologie

Informations

• Publié par: johannes_gutenberg-universitat_mainz
• Publié le: 01 janvier 2008
• Nombre de lectures: 32
• Langue: English
• Poids de l'ouvrage: 21 Mo

Extrait

Patterning of Tethered Bilayer Lipid
Membranes





Dissertation zur Erlangung des Grades
“Doktor der Naturwissenschaften”
im Promotionsfach Chemie


am Fachbereich Chemie, Pharmazie und Geowissenschaften
der Johannes Gutenberg – Universität Mainz







Mathieu Jung
Geboren in Straßburg, Frankreich

Mainz, 2008






















“One does not discover new lands without consenting to lose sight of the shore for a very
long time.”

André Gide






Table of contents

1. Introduction .................................................................................................................. 1
1.1. Construction and function of biomembranes ......................................................... 1
1.2. Model membranes .................................................................................................. 3
1.3. Patterning of bilayers ............................................................................................. 5
1.4. Topochemical reactions.......................................................................................... 6
1.4.1. Definition ....................................................................................................... 6
1.4.2. The concept of “reaction cavity”.................................................................... 7
1.5. Aim of this work: ................................................................................................... 8
2. Materials & Methods ................................................................................................. 10
2.1. Nuclear Magnetic Resonance (NMR) .................................................................. 10
2.2. Mass spectrometry................................................................................................ 11
2.3. Differential scanning calorimetry......................................................................... 11
2.4. Langmuir – Blodgett films ................................................................................... 12
2.4.1. Isotherms ...................................................................................................... 12
2.4.2. Film deposition............................................................................................. 16
2.5. Membrane Preparation ......................................................................................... 17
2.5.1. Template stripped gold (TSG)...................................................................... 18
2.5.2. Cleaning of oxide surfaces ........................................................................... 19
2.5.3. Monolayer formation by self-assembly........................................................ 19
2.5.4. Bilayer formation by vesicle fusion ............................................................. 20
2.5.5. Bilayer formation by rapid solvent exchange method.................................. 21
2.6. Water contact angle (WCA)................................................................................. 22
2.7. Ultraviolet-Visible Spectroscopy (UV-Vis)......................................................... 23
2.8. Electrical Impedance Spectroscopy (EIS)............................................................ 24
2.8.1. Physical basis: definition of impedance ....................................................... 24
2.8.2. Measurement setup....................................................................................... 26
2.8.3. Evaluation of the impedance data ................................................................ 27
2.9. Optical waveguide spectroscopy.......................................................................... 28
2.10. Fluorescence recovery after photobleaching (FRAP) ...................................... 29
2.10.1. Measurement procedure and evaluation....................................................... 30
2.10.2. Theory of diffusion....................................................................................... 32
3. Ultra flat heterogeneously structured substrates .................................................... 34
3.1. Photolithographic Structuring .............................................................................. 34
3.1.1. Concept......................................................................................................... 34
3.1.2. Photolithography .......................................................................................... 35
3.1.3. Success and limits of the method ................................................................. 36
3.2. Direct evaporation through a mask ...................................................................... 39
3.2.1. Protection of the TSGs with monolayers ..................................................... 39
3.2.2. Direct evaporation ........................................................................................ 40
3.2.3. Selective functionalization of the substrates ................................................ 43
4. Synthesis...................................................................................................................... 46
4.1. General remarks ................................................................................................... 46
4.2. Ether linked molecules ......................................................................................... 47
4.2.1. General procedure ........................................................................................ 47
4.2.2. Synthesis of the spacer part.......................................................................... 53
4.2.3. Discussion of the synthesis way................................................................... 54
4.3. Ester linked molecules ......................................................................................... 57
4.3.1. Double stranded anchor lipids...................................................................... 57
4.3.2. Single stranded anchor lipids ....................................................................... 62
5. Thermotropic behavior of polymerizable tBLM..................................................... 66
5.1. DSC experiments.................................................................................................. 66
5.2. Optical waveguide spectroscopy.......................................................................... 68
6. Langmuir films ........................................................................................................... 70
6.1. TD2(EO) LA and TD2(EO) TES ........................................................................ 71 4 4
6.2. HC1(EO) LA and HC1(EO) TES........................................................................ 73 4 4
6.3. HC2(EO) LA and HC2(EO) TES........................................................................ 74 4 4
7. UV-Visible spectroscopy ............................................................................................ 76
7.1. HC2(EO) TES on quartz...................................................................................... 76 4
7.2. HC2(EO) LA on quartz........................................................................................ 78 4
8. Photolithographic patterning.................................................................................... 81
8.1. HC2(EO) TES on quartz...................................................................................... 81 4
8.2. HC2(EO) LA on quartz........................................................................................ 85 4
9. Electrical impedance spectroscopy ........................................................................... 88
9.1. Valinomycin ......................................................................................................... 88
9.2. Building the bilayer.............................................................................................. 89
9.3. Introduction of Valinomycin in the tBLM ........................................................... 90
9.4. Polymerization of the bilayer ............................................................................... 92
10. Fluorescence recovery after photobleaching ....................................................... 94
10.1. Sample preparation........................................................................................... 95
10.2. Bilayer formation ............................................................................................. 97
10.3. FRAP measurements ........................................................................................ 98
10.3.1. Lipid diffusion in both leaflets ..................................................................... 98
10.3.2. Lipid diffusion in the bottom leaflet .......................................................... 100
11. Conclusions and outlook...................................................................................... 101
12. Appendix ............................................................................................................... 105
112.1. H-NMR and FD-Mass spectra