Single Molecules and Nanotechnology

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Rudolf Rigler, H. Vogel
Springer Science & Business Media, Dec 7, 2007 - Technology & Engineering - 318 pages
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The investigation of molecules as individuals has grown rapidly in recent years, and in the process has uncovered molecular properties not normally accessible by ensemble experiments. In particular, the direct characterization of biologically important molecules such as enzymes, molecular motors, or receptors and entire signaling complexes in action, for example in a live biological cell, yielded un- pected insights. Common approaches for studying single molecules include the electrical detection of ion channels in membranes, the measurement of the dynamics of (bio)chemical reactions between individual molecules, the imaging of individual molecules by scanning probe techniques or by fluorescence correlation spectr- copy, and the direct monitoring of single molecules by optical microscopies, to mention a few. The application of these techniques in physics, chemistry, and bi- ogy has opened new areas of nanotechnology. This book provides a representative selection of recent developments in the rapidly evolving field of single molecule techniques of importance in life sciences and will have future impact on the quan- tative description of biological processes. The editors of this book hope that the chapters, written by leading scientists in the field, will attract students and scientists from different disciplines, provide them with an authentic insight into this young field of research, allow them to evaluate experimental methods and results, and thereby give them support for their own research. Lausanne Rudolf Rigler September 2007 Horst Vogel v Contents 1 Nanophotonics and Single Molecules. . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 W. E. Moerner, P. James Schuck, David P.
 

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Contents

134 Local Enhancement of the Optical Fields Near the Nanoantenna
14
135 Exploring the Chemical Enhancement for SurfaceEnhanced Raman Scattering with Au Bowtie Nanoantennas
15
14 Summary and Prospects
19
References
20
III
24
21 Introduction
26
Success Stories and Limitations
28
214 SingleMolecule Optics Under Cryogenic Conditions
29
22 Spectroscopy of Single LightHarvesting Complexes
30
222 The Static Electronic Structure of LH2 from Rps acidophila
34
223 Dynamics in the B800 Band of LH2 from Rs molischianum
38
224 Outlook of LightHarvesting Complex Spectroscopy
42
24 Conclusions
47
References
48
IV
53
31 Introduction
54
Semiconductor Systems of Reduced Dimensionality
56
322 Exciton States Quantum Confinement and Quantization of Energy
57
323 Optical Properties of Quantum Dots
58
From Physical Chemistry to Bioimaging
60
332 Quantum Dots Bioconjugates for Imaging
61
From Ensemble to SingleMolecule Sensitivity
63
341 Techniques for SingleMolecule Fluorescence Visualization
64
342 SingleMolecule Sensitivity Requirements
65
Applications in SingleMolecule Optical Imaging
71
352 Quantum Dot Tracking of CellSurface Membrane Receptors
73
36 Future Directions
74
37 Concluding Remarks
75
VI
82
41 Introduction
84
42 Theory
85
423 Image Correlation Spectroscopy
87
425 Scanning Fluorescence Correlation Spectroscopy
88
43 Applications
94
44 Implementation
97
441 Light Source
98
443 Electronics
99
444 Software
100
445 Data analysis
101
45 Conclusion
102
References
103
VIII
107
51 SingleMolecule Fluorescence Microscopy in Living Cells
108
52 Model Organism for Studying Signal Transduction
110
53 Optical SetUp and Data Acquisition
111
54 Autofluorescence of Dictyostelium Discoideum Cells
112
55 Data Analysis
114
552 Identification of Single Molecules
116
56 Stoichiometry of Receptors in the Plasma Membrane
119
57 Membrane Organization
121
571 Fluorescent Lipid Insertion
122
573 cAR1 Mobility
123
58 Prospects
125
References
126
IX
130
61 Introduction
132
63 Techniques
133
632 SingleParticle Tracking
135
633 Advanced Fluorescence Labeling
139
634 Statistical Analysis
144
64 Investigations
148
74 Conclusions and Perspectives
177
References
178
XIV
181
81 Introduction
182
821 Current Questions in Protein Folding
183
83 SingleMolecule Spectroscopy
184
831 Free Diffusion Experiments
188
832 Experiments on Immobilized Proteins
190
84 SingleMolecule Protein Folding
195
842 Immobilized Protein Molecules
202
85 Perspective
207
References
208
XV
217
91 Introduction
218
922 Membrane Surface Properties Defined by Phospholipid Composition
221
923 Mechanical Properties of Phospholipid Membranes
222
924 SelfAssembly of Vesicular Systems
224
93 Fabrication Transformations and Modifications of NanotubeVesicle Networks
225
932 Complex Network Geometries Caused by SelfOrganization
227
933 Internal Functionalization Mixing and Compartmentalization of NanotubeVesicle Networks
228
94 Single Molecule Applications in NanotubeVesicle Networks
230
942 TensionControlled Marangoni Lipid Flow and Intratubular Liquid Flow in Nanotubes
232
943 Electrophoretic Transport
238
944 Diffusive Transport
242
95 Summary
244
References
246
XVIII
251
101 The Nanoreactor Approach to SingleMolecule Chemistry
252
102 Properties of the αHL Nanoreactor
254
103 Early Related Work
256
104 Irreversible SingleMolecule Covalent Chemistry
258
105 Reversible SingleMolecule Covalent Chemistry
259
106 Turnover of Irreversible Reactions in a TwoCompartment System
261
107 Examining a Polymerization One Step at a Time
262
108 SingleMolecule Experiments are Capable of Revealing Information that Would be Obscured in Ensemble Measurements
264
109 Alternative Approaches to SingleMolecule Chemistry
265
1010 Problematic Issues and Present Limitations of the Approach
269
1011 Future Directions
272
References
274
XIX
278
111 Introduction
280
112 HighResolution Imaging Using Atomic Force Microscopy
282
1122 AFM Imaging
283
1124 Determining Oligomeric State and Assembly
286
1125 Observing Native Membranes
287
1126 AFM a Structural Biology Method Complementing Biochemical Procedures
288
1128 Observing Structural Flexibility of Membrane Proteins
289
1129 Imaging Conformational Changes of Native Membrane Proteins
292
113 SingleMolecule Force Spectroscopy of Membrane Proteins
294
1131 Molecular Interactions Establish Stable Structural Segments Within Membrane Proteins
295
1132 Mapping Molecular Interactions Stabilizing Structural Segments
297
1133 Membrane Proteins Choosing Different Unfolding Pathways and Crossing Different Barriers
298
1134 ProteinProtein Interactions Influence Stability and Unfolding Pathways of Membrane Proteins
299
1136 Unfolding Pathways Depend on Temperature
300
1137 How Do Secondary Structure Elements Unfold at Zero Force?
301
1139 Determining Natural Unfolding Rates of Secondary Structure Elements
302
11311 Observing Folding Steps and Kinetics of Single Membrane Proteins
304
115 Conclusion and Perspectives
305
References
306
Index
313
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