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Fixation for Electron Microscopy
- 1st Edition - November 12, 2012
- Editor: M.A. (Eric) Hayat
- Language: English
- Paperback ISBN:9 7 8 - 0 - 1 2 - 4 1 4 4 1 1 - 8
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 1 4 7 2 9 - 3
Fixation for Electron Microscopy presents how to better understand the effects of fixatives on the molecular structure of the cell. This book attempts to consider each aspect of… Read more
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Request a sales quoteFixation for Electron Microscopy presents how to better understand the effects of fixatives on the molecular structure of the cell. This book attempts to consider each aspect of fixation, including chemical interactions between fixatives and individual cellular substances. The chemistry of fixative interactions that are discussed in the book is based primarily on the reactions of a fixative with isolated proteins, lipids, nucleic acids, and carbohydrates. The book shows that the correct interpretation of information retrieved from electron micrographs depends on the knowledge of the basic principles underlying the fixation procedure. Also, the book presents the fixation of both eukaryotic and prokaryotic specimens. The special fixation conditions for plant specimens are discussed in detail and have been allotted a whole chapter. Also emphasized in this book is the connection between morphology and biochemical aspects of preparatory treatments and the chemical basis of the formation of artifacts. This topic is useful in understanding the modifications of cell structures introduced during their processing. A guide for recognizing and minimizing major artifacts and fixation faults that are usually encountered is also presented in the book. This valuable resource will prove useful to both students and professionals in the field of biology and clinical medicine. Specimen preservation researchers can also benefit from this book.
Preface
1 Introduction
Text
2 Factors Affecting the Quality of Fixation
Tissue Specimen Size
Preparation of Tissue Blocks
Automatic Specimen Processing
Osmolarity
Ionic Composition of Fixative Solution
Vehicle Osmolality
Effects of Added Substances
Methods for Adjusting the Osmolarity
Recommended Osmolality
Measurement of Osmolality
Buffers
Buffer Types
Preparation of Buffers
pH
Fixative Penetration
Temperature of Fixation
Duration of Fixation
Fixative Concentration
3 Aldehydes
Glutaraldehyde (Glutaric Acid Dialdehyde)
Uses of Glutaraldehyde
Nature of Commercial Glutaraldehyde
Reaction with Proteins
Reaction with Lipids
Reaction with Nucleic Acids
Reaction with Carbohydrates
Effects of Aldehydes on Tissue Physiology
Osmolality
Osmolality of Glutaraldehyde
Temperature
Concentration
pH
Fixation Method with Glutaraldehyde at Higher pHs
Rate of Penetration
Shrinkage Caused by Aldehydes
Heated Glutaraldehyde
Limitations of Glutaraldehyde
Purification of Glutaraldehyde
Determination of Glutaraldehyde Concentration
Storage of Glutaraldehyde
Glutaraldehyde-Containing Fixatives
Formaldehyde
Reaction with Proteins
Reaction with Lipids
Reaction with Nucleic Acids
Reaction with Carbohydrates
Preparation of Paraformaldehyde
Acrolein
Reaction with Proteins
Reaction with Lipids
Precaution in the Handling of Acrolein
Acrolein Fixatives
Polyaldehyde
Role of Aldehydes in Quantitative Electron Microscopy
Tissue Storage
4 Osmium Tetroxide
Introduction
Reaction with Unsaturated Lipids
Reaction with Saturated Lipids
Reaction with Proteins
Reaction with Lipoproteins
Reaction with Nucleic Acids
Reaction with Carbohydrates
Reaction with Phenolic Compounds
Loss of Lipids
Loss of Proteins
Swelling Caused by Osmium Tetroxide
Parameters of Fixation
Concentration of Osmium Tetroxide
Temperature of Fixation
Rate of Penetration
Duration of Fixation
Osmium Tetroxide as a Primary Fixative
Osmium Tetroxide as a Vapor Fixative
Removal of Bound Osmium from Sections
Osmium Blacks
Osmeth
Preparations for and Precautions in the Handling of Osmium
Tetroxide
Regeneration of Used Osmium Tetroxide
5 Permanganates
Potassium Permanganate Reactions
Reaction with Membranes
Reaction with Monoamines
Reaction with Other Cellular Substances
Contrast and Loss of Cellular Substances
Comparison between Potassium Permanganate and Osmium
Tetroxide
Fixation Procedures
Permanganate Fixatives
6 Miscellaneous Fixatives
Ruthenium Tetroxide
Dimethylsuberimidate
Carbodiimides
Trioxsalen
7 Methods of Fixation
Double Fixation
Glutaraldehyde and Osmium Tetroxide Mixture
Simultaneous Fixation for Light and Electron Microscopy
Anhydrous Fixation
8 Modes of Fixation
Vascular Perfusion
Considerations in Achieving Satisfactory Vascular Perfusion
Apparatus for Vascular Perfusion
Methods of Vascular Perfusion
Immersion Fixation
Dripping Method
Injection Method
Anesthesia
9 Effects of Fixation
Effect of Fixation on Actin
Effect of Fixation on Brain Tissue
Effect of Fixation on Kidney Tissue
Renal Medulla
Biopsy Specimens
Effect of Fixation on Membrane Fracture
Effect of Fixation on Mesosomes
Effect of Fixation on Microtubules and Microfilaments
Effect of Fixation on Mitochondria
Effect of Fixation on Myelin
Effect of Fixation on Plant Virus
Effect of Fixation on Plasma Membrane
Effect of Fixation on Staining
Effect of Fixation on Synaptic Vesicles
Effect of Fixation on Tight Junctions
Effect of Fixation on Specimens for X-Ray Microanalysis
Effect of Fixation on Vascular Endothelium
Artifacts
10 Changes in Specimen Volume
Text
11 Postmortem Changes
Text
12 Plant Specimens
Text
13 Fixation for Scanning Electron Microscopy
Cleaning of Specimens
Preservation of Microbial Association to Intestinal Epithelium
Relaxation Procedures
Fixation Process
Parameters of Fixation
Mode of Fixation
Specimen Size
Temperature
Duration of Fixation
Specimen Shrinkage
Fixatives
14 Fixation for Enzyme Cytochemistry
Comparison of Different Aldehydes
Effect of Aldehydes on Enzymes
Factors Affecting the Preservation of Enzyme Activity
Specimen Size
Concentration of Aldehyde
Duration of Fixation
Temperature
pH
Buffers
Mode of Fixation and Incubation
Single-Cell Specimens
Subcellular Fractions
Cryoultramicrotomy
15 Fixation for Immunoelectron Microscopy
Fixatives
16 Use of Dimethylsulfoxide in Fixation
Effect on Enzymes
Effect on Ultrastructure
17 Criteria for Satisfactory Specimen Preservation
Text
18 Artifacts
Transmission Electron Microscopy
Scanning Electron Microscopy
19 Specific Fixation Methods
Text
Appendix
Balanced Salt Solutions
Commonly Used Salts and Their Physicochemical Properties
Commonly Used Chemicals
References
Index
- No. of pages: 521
- Language: English
- Edition: 1
- Published: November 12, 2012
- Imprint: Academic Press
- Paperback ISBN: 9780124144118
- eBook ISBN: 9780323147293
MH
M.A. (Eric) Hayat
Dr. Hayat serves as Distinguished Professor in the Department of Biological Sciences at Kean University. He is an internationally renowned scholar in the fields of electron microscopy and cancer research, has authored and edited more than 31 books, several of these recently with Elsevier (dealing with immunohistochemistry of carcinomas, cancer imaging, and autophagy).
Dr. Hayat has published chapters in 54 books, and authored 10 of them. Seven major publishers have published his books: Cambridge University Press, Elsevier, Academic Press, McGraw-Hill, Plenum Press, CRC Press, and Van Nostrand Reinhold Company. Dr. Hayat’s books have been published in Japanese. He has also published a large number of articles in scientific journals and established the Electron Microscopy Center at Kean University.
Degree Information: Ph.D., Biology, Indiana University
Courses Taught: Electron Microscopy; Cell Biology; Seminar
Primary Area of Expertise: Electron microscopy, cell biology, cancer, health and disease. His research interests include immunohistochemistry and in situ hybridization of human carcinomas, including brain metastases and various primary cancers (lung, breast, prostate, colorectal, pancreatic, ovarian, gastrointestinal, and liver).
Advice For Students Preparing for Your Class: "Read the books and work very hard."
Affiliations and expertise
Department of Biological Science, Kean University, USARead Fixation for Electron Microscopy on ScienceDirect