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| 1 | High-Resolution Optical and Confocal Microscopy | 1 |
| Olaf Hollricher and Wolfram Ibach | ||
| 1.1 | Introduction | 1 |
| 1.2 Introduction to Theoretical Considerations in High-Resolution | ||
| Microscopy | 3 | |
| 1.3 | Introduction to Confocal Microscopy | 3 |
| 1.4 | Electromagnetic Scattering in Optical Systems | 4 |
| 1.5 | 3D-Intensity Distribution in the Focus | 5 |
| 1.5.1 | Large Aperture Angles | 6 |
| 1.5.2 | Transition to Small Aperture Angles | 9 |
| 1.6 | Theory of Image Formation | 11 |
| 1.6.1 | Microscope | 11 |
| 1.6.2 | Confocal Microscope | 12 |
| 1.6.3 | Confocal Raman Microscope | 12 |
| 1.7 | Image Formation for Light Scattering | 13 |
| 1.7.1 | Scattering Point | 13 |
| 1.7.2 | Reflection at a Mirror | 15 |
| 1.8 | Image Formation for Raman Scattering | 16 |
| 1.8.1 | Raman Emitting Point | 16 |
| 1.8.2 | Raman Emission of a Layer | 17 |
| 1.9 | Pinhole Size | 18 |
| References | 20 | |
| 2 | Introduction to the Fundamentals of Raman Spectroscopy | 21 |
| Benjamin Dietzek, Dana Cialla, Michael Schmitt, and Jürgen Popp | ||
| 2.1 | Introduction | 21 |
| 2.2 | Classical Picture of Light Scattering | 22 |
| 2.2.1 | Frequency Dependence of Raman Scattering | 22 |
| 2.2.2 Classical "Selection Rule" and Comparison to IR | ||
| Absorption | 24 | |
| 2.2.3 | Scattered Raman Intensity | 25 |
| 2.2.4 | Shortfalls of the Classical Picture | 27 |
| 2.3 | Raman Cross-Section Enhancement Mechanisms | 27 |
| 2.3.1 | Resonant Raman Scattering | 28 |
| 2.3.2 Advantages and Applications of Resonance | ||
| Raman Scattering | 31 | |
| 2.3.3 | Surface-Enhanced Raman Scattering | 34 |
| References | 40 | |
| 3 | Raman Instrumentation for Confocal Raman Microscopy | 43 |
| Olaf Hollricher | ||
| 3.1 | Introduction | 43 |
| 3.2 | The Development of Raman Microscopes | 44 |
| 3.3 | Confocality | 45 |
| 3.4 | Throughput of a Confocal Raman Microscope | 46 |
| 3.4.1 | Laser Wavelength | 47 |
| 3.4.2 | Excitation Power | 47 |
| 3.4.3 | Objective | 48 |
| 3.4.4 | Microscope Throughput | 48 |
| 3.4.5 | Coupling Between Microscope and Spectrometer | 49 |
| 3.4.6 | Spectrometer Throughput | 49 |
| 3.4.7 | Gratings | 50 |
| 3.4.8 | CCD Detector | 51 |
| 3.5 | Conclusion | 60 |
| References | 60 | |
| 4 Software Requirements and Data Analysis in Confocal | ||
| Raman Microscopy | 61 | |
| Thomas Dieing and Wolfram Ibach | ||
| 4.1 | Introduction | 61 |
| 4.2 | Requirements for Data Acquisition Software | 62 |
| 4.2.1 | Data Acquisition | 62 |
| 4.2.2 | Correlation of Spatial and Spectral Data | 63 |
| 4.3 Description of the Data Sets Acquired in Confocal | ||
| Raman Microscopy | 65 | |
| 4.4 | Pre-processing of Raman Spectra | 66 |
| 4.4.1 | Cosmic Ray Removal | 66 |
| 4.4.2 | Smoothing | 68 |
| 4.4.3 Background Substraction and Subtraction | ||
| of Reference Spectra | 70 | |
| 4.5 | Image Generation | 72 |
| 4.5.1 | Univariate Image Generation | 72 |
| 4.5.2 | Multivariate Image Generation | 75 |
| 4.6 | Image Masks and Selective Average Calculation | 81 |
| 4.7 | Combination of Single Spectra with Multi-spectral Data Sets | 82 |
| 4.7.1 | Basis Spectra | 83 |
| 4.7.2 | Fitting Procedure | 83 |
| 4.8 | Combination of Various Images | 84 |
| 4.9 | The Law of Numbers | 85 |
| 4.10 | Materials and Methods | 88 |
| References | 89 | |
| 5 Nano-spectroscopy of Individual Carbon Nanotubes and Isolated | ||
| Graphene Sheets | 91 | |
| Alain Jungen | ||
| 5.1 | Introduction | 91 |
| 5.2 | Individual Carbon Nanotubes | 92 |
| 5.2.1 | Phonons | 92 |
| 5.2.2 | Theory | 95 |
| 5.2.3 | Experiment | 96 |
| 5.2.4 | Microscopy | 98 |
| 5.2.5 | Thermography | 100 |
| 5.3 | Isolated Graphene Sheets | 101 |
| 5.3.1 | Theory | 101 |
| 5.3.2 | Experiment | 102 |
| 5.3.3 | Charge Distributions | 105 |
| 5.4 | Conclusion | 107 |
| References | 108 | |
| 6 Raman Spectroscopy and Confocal Raman Imaging in Mineralogy | ||
| and Petrography...............................................Ill | ||
| Marc Fries and Andrew Steele | ||
| 6.1 Introduction..............................................Ill | ||
| 6.2 Raman Spectroscopy and Imaging as a Mineralogy/Petrography | ||
| Tool | 112 | |
| 6.2.1 | Working with Thin Sections | 114 |
| 6.2.2 | Control of Laser Power | 116 |
| 6.3 | "Raman Mineralogy" Using Imaging Raman Techniques | 119 |
| 6.3.1 | Mineral Phase Imaging | 120 |
| 6.3.2 | Crystallographic Orientation Imaging | 122 |
| 6.3.3 | Phase Composition Imaging | 123 |
| 6.4 | Examples of "Raman Petrography" Applications | 124 |
| 6.4.1 | Raman Analysis of Shocked Minerals | 124 |
| 6.4.2 | Contextual Imaging of Carbonaceous Materials | 126 |
| 6.4.3 | Fluid Inclusions | 129 |
| 6.4.4 | Ancient Terrestrial Carbonaceous Materials | 130 |
| 6.5 | Raman Mineralogy in Field Geology Studies | 130 |
| 6.5.1 | Extraterrestrial Exploration | 132 |
| 6.6 | Conclusion | 133 |
| References | 133 | |
| 7 Raman Micro-spectral Imaging of Cells and Intracellular Drug | ||
| Delivery Using Nanocarrier Systems | 137 | |
| Christian Matthaus, Tatyana Chernenko, Luis Quintero, Milos | ||
| Miljkovic, Lara Milane, Amit Kale, Mansoor Amiji, Vladimir | ||
| Torchilin, and Max Diem | ||
| 7.1 | Introduction | 137 |
| 7.2 | Method | 140 |
| 7.2.1 | Data Acquisition | 140 |
| 7.2.2 | Introduction to Data Processing Methods | 140 |
| 7.2.3 | Experimental | 144 |
| 7.3 | Results | 145 |
| 7.3.1 | Cell Imaging | 145 |
| 7.3.2 | Drug Delivery Systems | 152 |
| 7.4 | Discussion | 161 |
| References | 162 | |
| 8 | Confocal Raman Microscopy in Pharmaceutical Development | 165 |
| Thomas F. Haefele and Kurt Paulus | ||
| 8.1 | Introduction | 165 |
| 8.1.1 Vibrational Spectroscopy in Pharmaceutical | ||
| Development | 166 | |
| 8.1.2 | Imaging in Pharmaceutical Development | 169 |
| 8.1.3 | Chemical Imaging in Pharmaceutical Development | 172 |
| 8.1.4 Confocal Raman Microscopy in Pharmaceutical | ||
| Development | 176 | |
| 8.2 Applications of Confocal Raman Microscopy in Pharmaceutical | ||
| Development | 179 | |
| 8.2.1 | Practical Considerations | 179 |
| 8.2.2 Investigation of Solid Dosage Forms by Chemical | ||
| Imaging | 185 | |
| 8.3 | Conclusions | 198 |
| 8.4 | Materials and Methods | 198 |
| References | 199 | |
| 9 | Characterization of Therapeutic Coatings on Medical Devices | 203 |
| Klaus Wormuth | ||
| 9.1 | Background | 203 |
| 9.2 | Passive Therapeutic Coatings | 205 |
| 9.2.1 | Coating Thickness | 205 |
| 9.2.2 | Swelling of Hydrophilic Gel Coatings | 208 |
| 9.3 | Active Therapeutic Coatings | 210 |
| 9.3.1 | Coating Morphology | 210 |
| 9.3.2 Drug Mixed with Single Polymer: Morphology | ||
| andElution | 211 | |
| 9.3.3 Drug Mixed with Two Polymers: Blending | ||
| and Layering | 216 | |
| 9.3.4 | Drug Mixed with Two Polymers: Exposure to Water | 219 |
| 9.3.5 Drug Mixed with Biodegradable Polymer: Drug Elution | ||
| with Polymer Degradation | 220 | |
| 9.4 | Summary | 223 |
| References | 223 | |
| 10 | Raman Imaging of Plant Cell Walls | 225 |
| Notburga Gierlinger | ||
| 10.1 | Introduction | 225 |
| 10.2 | Plant Cell Walls | 226 |
| 10.3 | Micro-Raman Spectroscopy of Plant Fibers | 227 |
| 10.4 | Plant Cell Wall Imaging by Confocal Raman Microscopy | 228 |
| 10.4.1 Imaging Cellulose Orientation and Lignin Distribution | ||
| in Wooden Cell Walls | 228 | |
| 10.4.2 Silica and Cell Wall Composition in Horsetail | ||
| (E. hyemale) | 232 | |
| 10.5 | Outlook | 234 |
| 10.6 | Material and Methods | 234 |
| References | 235 | |
| 11 | Confocal Raman Imaging of Polymeric Materials | 237 |
| Ute Schmidt, Jörg Müller, and Joachim Koenen | ||
| 11.1 | Introduction | 237 |
| 11.2 | Raman Imaging of Isotactic Polypropylene (iPP) | 239 |
| 11.3 | Raman Imaging of Polymer Blends | 244 |
| 11.3.1 Raman Imaging of Thin Films of the Polymer Blend: | ||
| Polystyrene (PS)- Ethyl-Hexyl-Acrylate (EHA) | 245 | |
| 11.3.2 Raman Imaging of Thin Films of the Polymer | ||
| Blend Ethyl-Hexyl-Acrylate (EHA)-Styrene-Butadiene | ||
| Rubber (SBR) | 247 | |
| 11.3.3 Raman Imaging of Thin Films of the Polymer Blend | ||
| PS-EHA-SBR | 249 | |
| 11.4 | Polymer Coatings | 251 |
| 11.4.1 | Acrylic Paints | 251 |
| 11.4.2 | Adhesives | 253 |
| 11.5 | Additives in Polymer Matrices | 253 |
| 11.6 | Summary | 257 |
| References | 257 | |
| 12 | Stress Analysis by Means of Raman Microscopy | 259 |
| Thomas Wermelinger and Ralph Spolenak | ||
| 12.1 | Introduction | 259 |
| 12.1.1 | Theoretical B ackground | 260 |
| 12.1.2 Measuring in Conventional Back-Scattering | ||
| Configuration | 261 | |
| 12.1.3 | Off-Axis Raman Spectroscopy | 263 |
| 12.1.4 Stress Tensor Analysis in Back-Scattering | ||
| Raman Microscopy | 266 | |
| 12.2 | Case Studies | 268 |
| 12.2.1 | 3D Raman Spectroscopy Measurements | 268 |
| 12.2.2 | ZnO | 271 |
| 12.2.3 The Influence of Stress on the Peak Position | ||
| of Polymers | 273 | |
| 12.3 | Discussion | 276 |
| References | 277 | |
| Index | 279 | |
