Sample Preparation Handbook for Transmission Electron Microscopy

✍ Scribed by Ayache

• Publisher: Springer
• Year: 2010
• Tongue: English
• Leaves: 360
• Category: Library

✦ Synopsis

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 specimen 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.

✦ Table of Contents

Foreword
Preface to the English Edition
About the Authors
Contents
1 Techniques: General Introduction
Conclusion
2 Preliminary Preparation Techniques
Sawing
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.3.1 Annular Wheel Saw
1.3.2 Electrochemical Cutting
1.3.3 Acid Saw
1.3.4 Spark Erosion Cutting
1.4 Advantages
1.5 Limitations
1.6 Compatible Techniques
1.7 Risks
1.8 Conclusion
Ultrasonic Cutting
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.3 Variants
2.3.1 Hollow Bit
2.3.2 Punching (Stamping)
2.3.3 Chemical Cutting
2.3.4 Electrochemical Cutting
2.3.5 Electro-erosion Cutting
2.4 Advantages
2.5 Limitations
2.6 Compatible Techniques
2.7 Risks
2.8 Conclusion
Mechanical Polishing
3.1 Principle
3.2 Operating Mode
3.2.1 Equipment and Supplies
3.2.2 Procedure
3.3 Variants
3.3.1 Vibrating Plate
3.3.2 Tripod Polishing
3.3.3 Mechanical/Chemical Polishing
3.3.4 Electropolishing
3.3.5 Chemical Polishing
3.4 Advantages
3.5 Limitations
3.6 Compatible Techniques
3.7 Risks
3.8 Conclusion
Dimpling
4.1 Principle
4.2 Operating Mode
4.2.1 Equipment and Supplies
4.2.2 Procedure
4.3 Variants
4.4 Advantages
4.5 Limitations
4.6 Compatible Techniques
4.7 Risks
4.8 Conclusion
Electropolishing
5.1 Principle
5.2 Operating Mode
5.2.1 Equipment and Supplies
5.2.2 Procedure
5.3 Variants
5.4 Advantages
5.5 Limitations
5.6 Compatible Techniques
5.7 Risks
5.8 Conclusion
Chemical Polishing
6.1 Principle
6.2 Operating Mode
6.2.1 Equipment and Supplies
6.2.2 Procedure
6.3 Variants
6.4 Advantages
6.5 Limitations
6.6 Compatible Techniques
6.7 Risks
6.8 Conclusion
Sandwich Technique
7.1 Principle
7.2 Operating Mode
7.2.1 Equipment and Supplies
7.2.2 Procedure
7.3 Variants
7.3.1 Variant 1
7.3.2 Variant 2
7.4 Advantages
7.5 Limitations
7.6 Compatible Techniques
7.7 Risks
7.8 Conclusion
Embedding
8.1 Principle
8.2 Operating Mode
8.2.1 Equipment and Supplies
8.2.2 Procedure
8.3 Variants
8.3.1 Double Embedding Variant
8.3.2 Extraction Variant
8.3.3 Variant in Specific Media
8.4 Advantages
8.5 Limitations
8.6 Compatible Techniques
8.7 Risks
8.8 Conclusion
SubstitutionInfiltrationEmbedding at Room Temperature
9.1 Principle
9.2 Operating Mode
9.2.1 Equipment and Supplies
9.2.2 Procedure
9.3 Variants
9.3.1 Variant 1
9.3.2 Variant 2
9.3.3 Variant 3
9.3.4 Variant 4
9.4 Advantages
9.5 Limitations
9.6 Compatible Techniques
9.7 Risks
9.8 Conclusion
SubstitutionInfiltrationEmbedding at Low Temperatures
10.1 Principle
10.2 Operating Mode
10.2.1 Equipment and Supplies
10.2.2 Procedure
10.3 Variants
10.3.1 Variant 1
10.3.2 Variant 2
10.3.3 Variant 3
10.3.4 Variant 4
10.4 Advantages
10.5 Limitations
10.6 Compatible Techniques
10.7 Risks
10.8 Conclusion
Appendix: Physical Preparation Protocols
Chemical Fixation
11.1 Principle
11.2 Operating Mode
11.2.1 Equipment and Supplies
11.2.2 Procedure
11.3 Variants
11.3.1 Fixation for the Immunolabeling and Cytochemical Techniques
11.3.2 Fixation of Materials Containing Air
11.3.3 Fixation of Materials by Perfusion
11.4 Advantages
11.5 Limitations
11.5.1 Spatial Conformation Changes
11.5.2 Loss of Chemical Elements
11.5.3 Structural Changes
11.5.4 Structural Transformation: Artifacts Due to the Tissue-Sampling Method
11.5.5 Chemical Changes: Artifacts Linked to the Composition of the Fixative Mixture
11.6 Compatible Techniques
11.7 Risks
11.8 Conclusion
Physical Fixation: Cryo-fixation
12.1 Principle
12.2 Operating Mode
12.2.1 Equipment and Supplies
12.2.2 Procedure
12.3 Variants
12.4 Advantages
12.5 Limitations
12.6 Compatible Techniques
12.7 Risks
12.8 Conclusion
Continuous Support Films
13.1 Principle
13.2 Operating Mode
13.2.1 Equipment and Supplies
13.2.2 Procedure
13.3 Variant
13.3.1 Ionization of Carbon Films
13.4 Advantages
13.5 Limitations
13.6 Compatible Techniques
13.7 Risks
13.8 Conclusion
Holey Support Films
14.1 Principle
14.2 Operating Mode
14.2.1 Equipment and Supplies
14.2.2 Procedure
14.3 Variant
14.4 Advantages
14.5 Limitations
14.6 Compatible Techniques
14.7 Risks
14.8 Conclusion
Bibliography
3 Thinning Preparation Techniques
Twin-Jet Electrolytic Thinning
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.4 Advantages
1.5 Limitations
1.6 Artifacts
1.7 Type of Analysis
1.8 Risks
1.9 Conclusion
Full-Bath Electrolytic Thinning (Window Technique)
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.3 Variants
2.3.1 Thinning a Single Side
2.3.2 Thinning a Disk
2.4 Advantages
2.5 Limitations
2.6 Artifacts
2.7 Type of Analysis
2.8 Risks
2.9 Conclusion
Twin-Jet Chemical Thinning
3.1 Principle
3.2 Operating Mode
3.2.1 Equipment and Supplies
3.2.2 Procedure
3.3 Variants
3.4 Advantages
3.5 Limitations
3.6 Artifacts
3.7 Type of Analysis
3.8 Risks
3.9 Conclusion
Full-Bath Chemical Thinning (Window Technique)
4.1 Principle
4.2 Operating Mode
4.2.1 Equipment and Supplies
4.2.2 Procedure
4.3 Variants
4.4 Advantages
4.5 Limitations
4.6 Artifacts
4.7 Type of Analysis
4.8 Risks
4.9 Conclusion
Ion Milling
5.1 Principle
5.2 Operating Mode
5.2.1 Equipment and Supplies
5.2.2 Procedure
5.3 Variants
5.4 Advantages
5.5 Limitations
5.6 Artifacts
5.7 Type of Analysis
5.8 Risks
5.9 Conclusion
Focused Ion Beam Thinning (FIB)
6.1 Principle
6.2 Operating Mode
6.2.1 Equipment and Supplies
6.2.2 Procedure
6.3 Variants
6.4 Advantages
6.5 Limitations
6.6 Artifacts
6.7 Type of Analysis
6.8 Risks
6.9 Conclusion
Bibliography
4 Mechanical Preparation Techniques
Crushing
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.3.1 Glass-Slide Technique
1.3.2 Cryo-crushing
1.3.3 Scraping Technique
1.4 Advantages
1.5 Limitations
1.6 Artifacts
1.7 Type of Analysis
1.8 Risks
1.9 Conclusion
Wedge Cleavage
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.3 Variants
2.3.1 Adapting the Method for the Small Angle Cleavage Technique (SACT)
2.4 Advantages
2.5 Limitations
2.6 Artifacts
2.7 Type of Analysis
2.8 Risks
2.9 Conclusion
Tripod Polishing
3.1 Principle
3.2 Operating Mode
3.2.1 Equipment and Supplies
3.2.2 Procedure
3.3 Variants
3.4 Advantages
3.5 Limitations
3.6 Artifacts
3.7 Type of Analysis
3.8 Risks
3.9 Conclusion
Ultramicrotomy
4.1 Principle
4.2 Operating Mode
4.2.1 Equipment and Supplies
4.2.2 Procedure
4.2.3 Observations
4.3 Variants
4.4 Advantages
4.5 Limitations
4.6 Artifacts
4.6.1 Examples of Defects
4.7 Type of Analysis
4.8 Risks
4.9 Conclusion
Cryo-ultramicrotomy
5.1 Principle
5.2 Operating Mode
5.2.1 Equipment and Supplies
5.2.2 Procedure
5.2.3 Observations
5.3 Variants
5.3.1 Tokuyasu Method
5.4 Advantages
5.5 Limitations
5.6 Artifacts
5.7 Type of Analysis
5.8 Risks
5.9 Conclusion
Bibliography
5 Replica Techniques
Direct Replica Technique
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.3.1 Carbon Film Variant
1.3.2 Metallic Film Variant
1.4 Advantages
1.5 Limitations
1.6 Artifacts
1.7 Type of Analysis
1.8 Risks
1.9 Conclusion
Indirect Replica
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.3 Variants
2.4 Advantages
2.5 Limitations
2.6 Artifacts
2.7 Type of Analysis
2.8 Risks
2.9 Conclusion
Extractive Replica
3.1 Principle
3.2 Operating Mode
3.2.1 Equipment and Supplies
3.2.2 Procedure
3.3 Variants
3.3.1 Two-Step Carbon/Plastic Extraction Replica Technique
3.4 Advantages
3.5 Limitations
3.6 Artifacts
3.7 Type of Analysis
3.8 Risks
3.9 Conclusion
Freeze Fracture
4.1 Principle
4.2 Operating Mode
4.2.1 Equipment and Supplies
4.2.2 Procedure
4.2.3 Observations
4.3 Variants
4.4 Advantages
4.5 Limitations
4.6 Artifacts
4.7 Type of Analysis
4.8 Risks
4.9 Conclusion
Bibliography
6 Techniques Specific to Fine Particles
Dispersion of Fine Particle Materials
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.3.1 Dry Dispersion
1.3.2 Dispersion on the Surface of Water (Langmuir Film)
1.3.3 Aerosol Dispersion
1.3.4 ''Spin Coating'' Dispersion
1.3.5 Dispersion and Spreading Nucleic Acids (DNA, RNA, and Associated Proteins)
1.4 Advantages
1.5 Limitations
1.6 Artifacts
1.7 Type of Analysis
1.8 Risks
1.9 Conclusion
Frozen Hydrated Film of Single Particles
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.2.3 Observation in Cryo-microscopy
2.3 Variants
2.3.1 Cryo-plunger in a Controlled Atmosphere Chamber
2.3.2 Cryo-fixation of a Contrasted Sample
2.4 Advantages
2.5 Limitations
2.6 Artifacts
2.7 Type of Analysis
2.8 Risks
2.9 Conclusion
Bibliography
7 Contrast Enhancement and Labeling Techniques
Decoration Shadowing
1.1 Principle
1.2 Operating Mode
1.2.1 Equipment and Supplies
1.2.2 Procedure
1.3 Variants
1.3.1 Bidirectional Shadowing
1.3.2 Rotary Shadowing
1.4 Advantages
1.5 Limitations
1.6 Artifacts
1.7 Type of Analysis
1.8 Risks
1.9 Conclusion
Negative Staining
2.1 Principle
2.2 Operating Mode
2.2.1 Equipment and Supplies
2.2.2 Procedure
2.2.3 Observations
2.3 Variants
2.4 Advantages
2.5 Limitations
2.6 Artifacts
2.7 Type of Analysis
2.8 Risks
2.9 Conclusion
Positive Staining
3.1 Principle
3.2 Operating Mode
3.2.1 Equipment and Supplies
3.2.2 Procedure
3.3 Variants
3.3.1 Variant for Spread-Out Molecules
3.4 Advantages
3.5 Limitations
3.6 Artifacts
3.7 Type of Analysis
3.8 Risks
3.9 Conclusion
Immunolabeling
4.1 Principle
4.2 Operating Mode
4.2.1 Equipment and Supplies
4.2.2 Procedure
4.2.3 Observations
4.3 Variants
4.3.1 Double Labeling
4.3.2 Labeling Fine Particle Materials
4.3.3 Freeze-Fracture Labeling
4.4 Advantages
4.5 Limitations
4.6 Artifacts
4.7 Type of Analysis
4.8 Risks
4.9 Conclusion
Bibliography
Glossary
Photo Credits
Index