Sample Preparation Handbook for Transmission Electron Microscopy (eBook)

Methodology

Jeanne Ayache, Luc Beaunier, Jacqueline Boumendil, Gabrielle Ehret, Daniele Laub (Autoren)

eBook Download: PDF

2010 | 2010
XXIII, 250 Seiten
Springer New York (Verlag)
978-0-387-98182-6 (ISBN)

Infos

Successful transmission electron microscopy in all of its manifestations depends on the quality of the specimens examined. Biological specimen preparation protocols have usually been more rigorous and time consuming than those in the physical sciences. For this reason, there has been a wealth of scienti?c literature detailing speci?c preparation steps and numerous excellent books on the preparation of b- logical thin specimens. This does not mean to imply that physical science specimen preparation is trivial. For the most part, most physical science thin specimen pre- ration protocols can be executed in a matter of a few hours using straightforward steps. Over the years, there has been a steady stream of papers written on various aspects of preparing thin specimens from bulk materials. However, aside from s- eral seminal textbooks and a series of book compilations produced by the Material Research Society in the 1990s, no recent comprehensive books on thin spe- men preparation have appeared until this present work, ?rst in French and now in English. Everyone knows that the data needed to solve a problem quickly are more imp- tant than ever. A modern TEM laboratory with supporting SEMs, light microscopes, analytical spectrometers, computers, and specimen preparation equipment is an investment of several million US dollars. Fifty years ago, electropolishing, chemical polishing, and replication methods were the principal specimen preparation me- ods.

Inhalt

Foreword 6
Preface to the English Edition 10
About the Authors 13
Contents 15
Abbreviations 21
1 Methodology: General Introduction 22
2 Introduction to Materials 24
Introduction 24
1.1 Origin of Materials 24
1.2 Evolution of Materials 24
1.3 General Problems Presented by Microstructure Investigations 25
Classification of Materials and Properties 27
2.1 Types of Chemical Bonds: Atomic and Molecular 27
2.2 Type of Materials and Chemical Bonds 29
2.3 Chemical Bonds and Mechanical Properties 29
2.3.1 Mechanical Properties and Crystallinity 30
2.3.2 Rigidity: From Hard to Soft 31
2.3.3 Tensile Strength: Ductility--Brittleness 31
2.3.4 Mechanical Properties of Organic Materials and Glass Transition (Tg) 33
Microstructures in Materials Science 34
3.1 Problems to Be Solved in Materials Science 34
3.2 Materials Microstructures 35
3.3 Polymer Microstructures 39
3.4 Crystalline Defects and Properties of Materials 40
3.5 Solid-State Polymer Properties 44
Microstructures in Biological Materials 45
4.1 Problems to Be Solved in Biology 45
4.2 Singularity of Biological Materials: Importance of the Liquid Phase 47
4.3 Microstructure in Biology 48
4.4 Role of Structures on Functional Properties 51
Bibliography 51
3 The Different Observation Modes in Electron Microscopy (SEM, TEM, STEM) 53
Introduction 53
Signals Used for Electron Microscopy 53
2.1 Electron--Matter Interaction 53
2.2 Signals Used for Imaging 55
2.3 Signals Used for Chemical Analysis 56
2.4 Signals Used for Structure 57
2.4.1 Transmitted Electrons: Thin Samples with Thickness 100 nm 57
Microscopes and Observation Modes 57
3.1 Illumination Sources 57
3.1.1 Thermionic Sources 58
3.1.2 Field Emission Guns (FEGs) 58
3.2 Illumination Modes and Detection Limits 59
3.3 Microscope Resolutions and Analysis 59
3.3.1 Resolution Limit of the TEM 59
3.3.2 Spatial Resolution 60
The Different Types of Microscopes: SEM, TEM, and STEM 61
4.1 Scanning Electron Microscope (SEM) 61
4.2 Conventional Transmission Electron Microscope (CTEM) 61
4.3 Analytical TEM/STEM Microscope and ''Dedicated STEM'' 64
Different TEM Observation Modes 66
5.1 Origin of Contrast 66
5.1.1 Amplitude Contrast and Phase Contrast 67
5.2 Diffraction Contrast Imaging Modes in TEM and TEM/STEM 68
5.3 Chemical Contrast Imaging Modes in TEM and TEM/STEM 69
5.4 Spectroscopic Contrast Imaging Modes in TEM and TEM/STEM 70
5.5 EDS Chemical Analysis Methods in TEM and TEM/STEM 71
5.6 EELS Spectroscopic Analysis Modes in TEM and TEM/STEM 72
Conclusion and Information Assessment 72
Bibliography 74
4 Materials Problems and Approaches for TEM and TEM/STEM Analyses 76
Introduction 76
Analyses Conducted Prior to TEM Analyses 76
2.1 Macroscopic Characterization 78
2.2 Microscopic Characterization 78
2.3 Microscopic and Nanoscopic Characterization 79
Approach for Beginning the Investigation of a Material 80
Selection of the Type of TEM Analysis 82
Analysis of Topography 82
Structural Analysis in TEM 83
6.1 Morphology and Structure of Materials 83
6.2 Atomic Structure 87
Crystallographic Analysis 89
Analysis of Crystal Defects: 1D (Dislocations), 2D (Grain Boundaries and Interfaces), and 3D (Precipitates) 91
EDS Chemical Analysis and EELS Spectroscopic Analysis 92
9.1 Phase Identification and Distribution 92
9.2 Concentration Profiles and Interface Analysis 93
Structural Analyses Under Special Conditions 94
10.1 In Situ Analyses 94
10.1.1 At Room Temperature 94
10.1.2 At High Temperatures 94
10.1.3 At Low Temperatures 95
10.2 Cryomicroscopy 95
10.2.1 Structure of Isolated Particles from Biological Materials or Polymers 95
10.2.2 Structure of Bulk Frozen Samples 96
Study of Properties 96
11.1 Optical Properties 96
11.2 Electrical Properties 96
11.3 Electronic Properties 96
11.4 Magnetic Properties 97
11.5 Mechanical Properties 97
11.6 Chemical Properties 97
11.7 Functional Properties 97
Relationship Between Sample Thickness and Analysis Type in TEM and TEM/STEM 100
Assessment of TEM Analyses 100
5 Physical and Chemical Mechanisms of Preparation Techniques 101
Introduction 101
Mechanical Action 102
2.1 Principles of a Material's Mechanical Behavior 102
2.2 Abrasion Principle 103
2.2.1 Techniques Involving Cutting by Means of Mechanical Abrasion: Sawing and Grinding 104
2.2.2 Abrasive Techniques: Mechanical Polishing, Dimpling, and Tripod Polishing 104
2.3 Rupture Principles 105
2.3.1 Techniques Involving Fracture: Crushing, Wedge Cleavage, Ultramicrotomy, and Freeze Fracture 105
Chemical Action 108
3.1 Principle of Chemical and Electrochemical Dissolution 108
3.1.1 Techniques Involving Chemical and Electrochemical Dissolution 110
Ionic Action 111
4.1 Ionic Abrasion Principles 111
4.2 Techniques Involving Ion Abrasion 112
4.2.1 Ion Beam Thinning and Focused Ion Beam Thinning (FIB) 112
Actions Resulting in a State Change of Materials Containing an Aqueous Phase 116
5.1 Elimination of the Aqueous Phase 116
5.2 Freezing Principles 118
5.3 Principle of Substitution, Infiltration, and Embedding in Cryogenic Mode 120
5.4 Cryo-sublimation (or Freeze-Drying) Principle 121
Actions Resulting in a Change in Material Properties 121
6.1 Chemical Fixation Principles 122
6.1.1 Constancy of pH 123
6.1.2 Molar Concentration 124
6.1.3 Ionic Concentration 124
6.2 Dehydration Principles 126
6.3 Infiltration Principles 126
6.4 Embedding or Inclusion Principles 128
6.5 ''Positive-Staining'' Contrast Principles 129
Physical Actions Resulting in Deposition 130
7.1 Physical Deposition 130
7.2 Physics of the Coating Process 131
7.2.1 Nature of Chemical Elements Used as Sources 132
7.2.2 Different Methods of Particle Production 133
7.2.3 Vacuum 137
7.2.4 Substrate 137
7.3 Techniques Involving a Physical Deposition: Continuous or Holey Thin Film, Contrast Enhancement by Shadowing or Decoration, Replicas, and Freeze Fracture 137
7.3.1 Replica Techniques 138
7.3.2 Contrast Enhancement by Physical Coating: ''Negative-Staining'' Contrast 138
Bibliography 140
Mechanical Action 140
Chemical Action 140
Ionic Action 140
Actions Resulting in a State Change of Materials Containing an Aqueous Phase 140
Actions Resulting in a Change in Material Properties 141
Physical Actions Resulting in a Deposit 141
6 Artifacts in Transmission Electron Microscopy 142
Introduction 142
Preparation-Induced Artifacts 142
2.1 Mechanical Preparation-Induced Artifacts 144
2.1.1 Secondary Thermal Damage Induced During Mechanical Preparation 146
2.2 Ionic Preparation-Induced Artifacts 147
2.2.1 Secondary Thermal Damage Induced During Ionic Preparation 148
2.3 Chemical Preparation-Induced Artifacts 148
2.3.1 Changes Specific to Biological Materials 149
2.3.2 Secondary Thermal Damage Induced During Chemical Preparation 150
2.4 Physical Preparation-Induced Artifacts 151
2.4.1 Secondary Thermal Damage Induced During Physical Preparation 151
Artifacts Induced During TEM Observation 152
3.1 Artifacts Not Linked to Thermal Damages 152
3.2 Secondary Thermal Damage 154
Examples of Artifacts 154
4.1 Artifacts Induced by the Tripod Polishing Technique 154
4.2 Artifacts Induced by the Ultramicrotomy Technique 157
4.3 Artifacts Induced by the Freeze-Fracture Technique 164
4.4 Artifacts Induced by Ion Milling or FIB 165
4.5 Artifacts Induced by the Substitution--Infiltration--Embedding Technique 171
4.6 Artifacts Induced by Chemical Fixation 171
4.7 Artifacts Induced by the Extractive-Replica Technique 172
4.8 Artifacts Induced by the Shadowing Technique 173
4.9 Artifacts Induced by the ''Positive-Staining'' Contrast Technique 174
4.10 Artifacts Induced by the Cryofixation Technique 175
4.11 Artifacts Induced by the Fine Particle Dispersion Technique 176
4.12 Artifacts Induced by the Frozen-Hydrated-Film Technique 178
4.13 Artifacts Induced by the ''Negative-Staining'' Contrast Technique 180
4.14 Artifacts Induced by the Electron Beam 181
Summary Tables 185
Bibliography 186
7 Selection of Preparation Techniques Based on Material Problems and TEM Analyses 188
Introduction 188
Classification of Preparation Techniques 188
Characteristics of Preparation Techniques 189
Criteria Used to Select a Preparation Technique 190
Selection Criteria Based on Material Type 190
Selection Criteria Based on Material Organization 193
6.1 Bulk Materials 193
6.2 Single-Layer or Multilayer Materials 193
6.3 Fine Particles 194
Selection Criteria Based on Material Properties 194
7.1 Based on the Physical State of the Material 194
7.2 Based on the Chemical Phases in the Material 194
7.3 Based on the Electrical Properties of the Material 195
7.4 Based on the Mechanical Properties of the Material 195
7.4.1 Materials in Solid-State Physics 195
7.4.2 Soft-Ductile Materials 196
7.4.3 Hard-Resistant Materials 196
7.4.4 Materials of Intermediate Hardness and Ductility 196
7.4.5 Biological Materials 197
Selection Criteria Related to the Type of TEM Analysis 198
8.1 Preparation Techniques 199
Selection of the Orientation of the Sample Section 200
9.1 Microstructure Geometry 204
9.2 Defect Geometry 206
Selection Criteria Related to Artifacts Induced by the Preparation Technique 206
Adaptation of the Technique Based on Problems Related to Observation 208
11.1 Reducing Sample Thickness 208
11.2 Increasing Contrast 209
11.3 Reducing Charge Effects 209
11.4 Limitation of Strain Hardening 209
11.5 Removal of Surface Amorphization 209
11.6 Removal of Surface Contamination 209
11.7 Final Cleaning of the Thin Slice 210
Conclusion 210
Bibliography 214
8 Comparisons of Techniques 215
Introduction 215
Examples Using Fine Particle Materials 215
2.1 Comparison of Mechanical Preparations and Replicas 215
2.2 Comparison of ''Negative-Staining'' Contrast and Freeze-Fracture Techniques 218
2.3 Comparison of ''Negative-Staining'' and Decoration-Shadowing Contrast Techniques 219
2.4 Comparison of ''Positive-Staining'' and Decoration-Shadowing Contrast Techniques 222
Examples Using Bulk or Multilayer Materials 223
3.1 Comparison Between Different Mechanical Preparations 223
3.2 Comparison Between Mechanical Preparations and Ionic Preparations 225
3.3 Comparison Between Mechanical Preparations and Electrolytic Preparations 236
3.4 Comparison Between Techniques Specific to Biology 238
3.5 Comparison Between All Techniques That Can Be Used in Biology on One Example: Collagen 245
Bibliography 249
9 Conclusion: What Is a Good Sample? 251
Photo Credits 253
Index 255

Produkt-Details

• Erscheint lt. Verlag: 3.7.2010
• Zusatzinfo: XXIII, 250 p.
• Verlagsort: New York
• Sprache: englisch
• Themenwelt: Naturwissenschaften ► Chemie
• Themenwelt: Technik ► Maschinenbau
• Schlagworte: Biological Microscopy • Biology • Chemical Analysis • electron microscopy • Materials Science • Microscopy • sample preparation physics • Sample preparation technique for TEM • spectroscopic analysis • structure analysis • thin slices protocols • Transmission Electron Microscopy
• ISBN-10: 0-387-98182-9 / 0387981829
• ISBN-13: 978-0-387-98182-6 / 9780387981826