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Histological Techniques for Electron Microscopy - 2nd Edition - ISBN: 9781483231921, 9781483263687

Histological Techniques for Electron Microscopy

2nd Edition

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Author: Daniel C. Pease
eBook ISBN: 9781483263687
Imprint: Academic Press
Published Date: 1st January 1964
Page Count: 398
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Histological Techniques for Electron Microscopy, Second Edition, offers a practical guide for those who would study cells or tissues with an electron microscope. The book contains 11 chapters and begins with a discussion of the organization and management of an electron microscope laboratory. This is followed by separate chapters on tissue preservation; fixatives and their variations; methacrylate embedding and cross-linked plastics, microtomes and microtomy; and section mounting. Subsequent chapters deal with the techniques of "staining" for electron microscopy, photography; mounting, shadowing, and replication; and negative staining.

Table of Contents

Preface to the First Edition

Preface to the Second Edition

1. Organization and Management of an Electron Microscope Laboratory

1.1. Laboratory Layout

1.2. Equipment

1.3. Management of the Laboratory

1.4. Service Personnel

1.5. Technical Help

l.6. Shared Facilities

1.7. Educating Administrators and Clinicians

2. Tissue Exposure

2.1. Introductory Remarks

2.2. Post-Mortem Change

2.3. Minced Tissue

2.4. Topical, In Situ, Preservation of Natural Surfaces

2.5. In Situ Fixation of Cut Surfaces

2.6. Vascular Perfusion

2.7. Hyaluronidase Pretreatment

2.8. Special Problems with Human Tissues

2.9. Special Problems with Tissue Cultures and Cell Suspensions

2.10. The Use of Cold-Blooded and Young Animals

2.11. Botanical Material

Literature Cited

3 . Fixation

3.1. Introductory Remarks

3.2. Palade (1952) Fixative, Buffered Osmium Tetroxide

3.3. Variations of Osmium Tetroxide Fixatives

3.4. Special Purpose Osmium Tetroxide Fixatives

3.5. Temperature Control of Osmium Tetroxide Fixation

3.6. Duration of Osmium Tetroxide Fixation

3.7. Criteria of Good Osmium Tetroxide Fixation

3.8. Aldehyde Fixation: Introductory Remarks

3.9. Buffered Aldehydes as Primary Fixatives

3.10. Aldehydes as Fixatives for Histochemistry

3.11. Morphological Features of Aldehyde Fixation

3.12. The Permanganate Fixative of Luft (1956)

3.13. Rapid Freezing

3.14. Freeze-Drying

3.15. Freeze-Substitution

Literature Cited

4. Embedding

4.1. Introductory Remarks

4.2. Dehydration

4.3. Transitional Solvents

4.4. Infiltration

4.5. Handling Thick Embedding Media

4.6. Desirable Characteristics of the Embedment

4.7. Methacrylate Embedding

4.8. "Polymerization Damage" and the Use of "Prepolymerized" Methacrylate

4.9. Limitations of Methacrylate

4.10. Araldite Embedding

4.11. Epon Embedding

4.12. Maraglas Embedding

4.13. Polyester Vestopale Embedding

4.14. Selectron, Rigolac and Viapale Embedding

4.15. Cross-linked Methacrylate Embedding

4.16. Water-soluble Resin Embedding

4.17. Mixed Plastic Embedding

4.18. Gelatin Embedding

4.19. Preparing the Block for Sectioning

Literature Cited

5. Sectioning

5.1. Introductory Remarks

5.2. The Original Porter-Blum Microtome

5.3. Modification of the Porter-Blum Microtome

5.4. LKB "Ultrotome"

5.5. Leitz Ultramicrotome

5.6. Reichert Ultramicrotome

5.7. Cambridge Ultramicrotome

5.8. SI-RO-FLEX Ultramicrotome

5.9. JUM-5 Ultramicrotome

5.10. Sorvall MT-2 Ultramicrotome

5.11. Clevenger's (1963) Microtome

5.12. Knives Used for Ultrathin Sectionin

5.13. Glass Knife Manufacture

5.14. Knife Angle

5.15. Examination and Selection of Glass Knives

5.16. Diamond Knives

5.17. Trough Construction

5.18. Mounting the Knife

5.19. Preferred Orientation of Block in Microtome

5.20. Section Flotation

5.21. Flattening Sections

5.22. Sectioning

5.23. Contamination—Principal Cause of Sectioning Difficulty!

5.24. Estimation of Section Thickness

5.25. Summary of Sectioning Technique

5.26. Picking Up Sections

Literature Cited

6. Section Mounting

6.1. Introductory Remarks

6.2. Parlodion Films

6.3. Formvar Films

6.4. Preparing Film Nets

6.5. The Mechanics of Carbon Evaporation

6.6. Carbon Stabilization of Naked Sections

6.7. Carbon Stabilization of Plastic Films

6.8. Pure Carbon Films Prepared on a Plastic Substrate

6.9. Stripping Pure Carbon Films

6.10. "Sandwiched" Sections

6.11. Serial Sections

6.12. Mounting Screens in the Microscope Specimen Holder

6.13. Dirty Specimens

Literature Cited

7. "Staining"

7.1. Introductory Remarks

7.2. Penetration of Stains

7.3. Alkaline Lead Hydroxide Stains

7.4. Uranyl Stains

7.5. Phosphotungstic Acid Stain

7.6. Lawn's Permanganate Stain

7.7. Vanadium Salt Stains

7.8. Chromyl Chloride Stain

7.9. Stain Dependence upon Fixation

7.10. Histochemical Applications

7.11. Specific Antibody "Stains"

7.12. Autoradiography

7.13. Thick Sections for Conventional Microscopy

7.14. Removal of Cured Epoxy Resins

Literature Cited

8. Microscopy

8.1. Introductory Remarks

8.2. Objective Aperture Position and Size

8.3. Objective Aperture Alignment

8.4. Aperture Cleaning and Manufacture

8.5. Microscope Maintenance

8.6. Alignment, General Remarks

8.7. Objective Alignment

8.8. Alignment of the Illuminating System

8.9. Source Alignment

8.10. Condenser Alignment

8.11. Filament Replacement and Alignment

8.12. Objective Lens Compensation

8.13. Instrument Stability

8.14. Trouble Shooting

General References

9. Photography

9.1. Introductory Remark

9.2. Section Thickness versus Magnification

9.3. Focus

9.4. Special Considerations in Picture Taking

9.5. Reasons for Unsatisfactory Pictures

9.6. Photographic Plates

9.7. Plate Development

9.8. Enlarging

9.9. Timing Print Exposures

9.10. "Dodging"

9.11. Problem Negatives, Reduction and Intensificaion

9.12. Interpretation of Electron Microscope Images

General Reference

10. Particulate Specimens , Mounting, Shadowing, and Replication

10.1. Introductory Remarks

10.2. Suspended and Fragmented Material

10.3. Shadow-Casting

10.4. Replication

10.5. Single-Stage Replication

10.6. Preshadowed Replicas

10.7. Two-Stage Replication

10.8. Preparation of Surfaces for Replication

10.9. Materials for Replication, Deposited from Solution or Melt

10.10. Materials for Replication, Deposited by Vacuum Evaporation

Literature Cited

General References

11. Particulate Specimens, Negative Staining

11.1. Introductory Remarks

11.2. Theoretical Aspects of Negative Staining

11.3. Negative Stains

11.4. Specimen Preparation for Negative Staining

11.5. Microscopy of Negatively Stained Material

Literature Cited

Appendix A. Outline of Basic Technique

Training Program

Appendix B. The Literature of Electron Microscopy


Published Records of International Meetings

Published Records of Major National Meetings

Appendix C. Some Sources of Equipment and Materials

General Sources

Specimen Grids


Calibration Supplies

Author Index

Subject Index


No. of pages:
© Academic Press 1964
1st January 1964
Academic Press
eBook ISBN:

About the Author

Daniel C. Pease

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