
Handbook of Microscopy
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Handbook of Microscopy is a manual that deals mainly with the basic instruments and techniques used in light microscopy and its biological applications. A large section is devoted to the study of organic matter in microfossils preserved in rocks, in view of its stratigraphic importance in mining and oil prospecting. This text is comprised of six chapters; the first of which introduces the reader to the basic principles as well as to the instruments and techniques used in light microscopy. This book also discusses the microscopes and electronic flashlights for photomicrography, along with the use of monochromatic light, stereological and physicochemical microanalysis, microanalysis by electron microscopy, and microdetermination of physical values. Attention then turns to staining and impregnation and methods of fixation, examination, cutting, and mounting. The remaining chapters focus on the microscopy of topological stains and non-specific cytological stains, with emphasis on special methods used in animal and plant histology and protistology and mycological methods in pathology. This book is written specifically for microscopists.
Table of Contents
Preface
Acknowledgments
Abbreviations
1 Instruments and Techniques
1.1 Basic Principles
Light and Colors
Image Formation
Mechanical Conventions
Aberrations
Depth of Field
Flatness of Field
Microscopic Image in Wave Optics
Useful Magnification
Homogeneous Immersion of Objectives
Antireflection Treatment of Optical Surfaces
1.2 Basic Instruments
Mechanical Features of the Microscope
Use of the microscope
Microscopic Observation
1.3 Chief Instrumental Methods
Dark Ground Illumination
Examination under Reflected Light
Infrared Microscopy
Ultraviolet Microscopy
Pupillary Modulators
Interference Microscopes
Phase Contrast
Polarizing Microscope
Study of Anisotropy of Biological Tissues under Polarized Light
Fluorescence Microscopy
Projection Microscopy
1.4 Recording of Images
Photomicrography
Photomacrography
Illuminating during Photomicrography
Colored Filters
Color Differences in Emission and Reception
Black and White Photography
Procedures for Color Photography
Balanced Color Compensation
Variations and Deficiencies in Color Emulsions
Emulsions
Exposure Time
Measurement of the Exposure Time
Development
Printing on Paper
Classification
Transposition of Contrasts
Special Effects in Photomicrography
Equidensitometry
1.5 Synthesis
Stereoscopic Photomicrographs
Graphic Reconstructions
Solid Reconstructions
Cine Reconstructions
Cinemicrography
Drawing at the Microscope
Artifacts
1.6 Measurements
Measuring Techniques
1.7 Special Techniques
Locating Objects
Special Stages and Warmed or Cooled Stages
Microscopic Observations under Increased Pressure
Automation in Microscopy
Microsurgery
1.8 Use of Monochromatic Light
Monochromators
Spectrographs
Spark Microspectroscopes and Emission Spectrography
Microphotometers and Microspectrophotometers
1.9 Stereological Microanalysis
Counting
Morphometry and Stereology
Microquantimeters
1.10 Physicochemical Microanalysis
Micro-Electrophoresis Cell
Thermal Micromethods
Microrefractometry
Autoradiography
Historadiography
Spodography
1.11 Microanalysis by Electron Microscopy
The Scanning Electron Microscope
The Transmission Electron Microscope
Ultra High-Voltage Electron Microscopy
Ultraviolet Excited Electron Microscopy
Proton Microscopy
Ion Microscopy
1.12 Microdetermination of Physical Values
Microdetermination of Osmotic Pressure
Differential Separation Techniques
Ultracentrifugation of Cells
Determination of Dry Weight of Cellular Constituents
Determination of Constituents by Specific Absorption
2 Methods of Fixation, Examination, Cutting and Mounting
2.1 Methods of Preparation
Test or Reference Preparations
Rehydration of Dried Material
Preservation Mixtures
2.2 Methods of Fixation
The Aims of Fixation
Accuracy of Preservation in Fixation
Simple Fixatives
Fixative Mixtures
Time for Fixation
Cytological Fixatives
Embedding Compatibility
Penetration
Compatibility of Stains
Decalcification
Quantity of Fixative
Choice Based on the Specimen
Stopping Fixation
Fixatives for Electron Microscopy
2.3 Methods of Bleaching
Demineralization and Softening
Softening of Keratin and Chitin
Maceration
Depigmentation
2.4 Observations of Living Specimens
Examination of Living Specimens
Vital Staining
Methods of Preparation for Examination of Microorganisms In Vivo
Equilibrated Media
Osmodialysis
2.5 Special Techniques
Thin Sections Produced by Polishing
Disintegration Techniques
Contrast-Enhancing Techniques
Drying and Dehydration
Dehydration in the Liquid Phase
Clearing Agents: Removal of Ethanol
Dehydrating Agents Which Can Be Used Immediately before Paraffin
Clearing Agents Which Can Be Used between 95% Ethanol and Paraffin
2.6 Embedding Methods
Non-Aqueous Media
Aqueous Media
Embedding in Paraffin
Embedding in Celloidin or Collodion
Embedding in Epoxy Resins, Methacrylate and Polyesters
Embedding in Hydrophilic Media
Embedding with Gel-Paraffin Mixtures
2.7 Sections
Microtomes
Cutting Tools
Automatic Honing Equipment
Cutting with a Microtome
Smoothing and Attachment of Sections
Removal of Paraffin before Staining
Ultrafine Sections for Transmission Electron Microscopy
Preparation of Supporting Membranes for Electron Microscopy
Fixation and Preservation of Small Specimens
2.8 Mounting of Preparations
Microscopic preparation
Media for Temporary observation
Permanent Mounting
Visibility Indices in the Mounting Media
Air-tightness of Cements and Varnishes
3 Stains
3.1 Principal Stains
Anthraquinones
Azines
Monoazides
Polyazides
Cyanines, Quinolines
Diazonium, Tetrazonium, Tetrazolium Salts
Indulins
Nitro-Stains
Oxazines
Pyrazolones
Quinone-Oximes
Phthalocyanins
Thiazines
Triarylmethanes
Xanthenes
Thiazoles
Polymethines
Mineral Dyes
Natural Pigments
Fluorochromes
3.2 Synonyms of the Most Important Stains
Glossary of Stains and Dyes
4 Staining and Impregnation
4.1 Methodology
General Considerations
Observations
Staining Materials
Modes of Action
Neutraistains
Staining Whole Pieces
Negative Stains
Infiltration and Injection
Automation of Staining
4.2 Applications
Non-specific Stains
5 Topological Stains
5.1 General Panoptic Methods
Staining of Nuclei with Progressive Haematein Lakes
Trichromes and Dichromes
5.2 Special Methods in Animal Histology
Staining of Blood and Intracellular Parasites
Staining of Bony and Cartilaginous Tissues
Staining of Muscle Fibers
Staining of the Nervous System
Bacteriological Stains
Staining of Tissues Containing Collagen
Staining of Glycogen and Galactogen
Staining of Integuments
Pigments
5.3 Special Methods in Plant Histology
Spores and Cysts
Plant Techniques
Detection of Parasitic Fungi in Plants
Mycological Techniques
5.4 Special Methods in Protistology
Techniques of Protistology
Amoeba
Sporozoa
Flagellates
Infusoria
Spirochaetae
5.5 Mycological Methods in Pathology
Zoopathological Techniques
5.6 Observations on Supplementary Techniques
Automation of Fixation, Impregnation and Staining
Large-Scale Series for Teaching
6 Non-specific Cytological Stains
6.1 General Cytology
6.2 Staining of Intracellular Organelles
6.3 Cellular Diagnosis in Cancer
6.4 Conclusions
Appendix I Physicochemical Constants
Bibliography
Index
Product details
- No. of pages: 334
- Language: English
- Copyright: © Butterworth-Heinemann 1983
- Published: January 19, 1983
- Imprint: Butterworth-Heinemann
- eBook ISBN: 9781483164878