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Fixation for Electron Microscopy - 1st Edition - ISBN: 9780123339201, 9780323147293

Fixation for Electron Microscopy

1st Edition

Editor: M Hayat
eBook ISBN: 9780323147293
Imprint: Academic Press
Published Date: 28th December 1981
Page Count: 521
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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 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.

Table of Contents


1 Introduction


2 Factors Affecting the Quality of Fixation

Tissue Specimen Size

Preparation of Tissue Blocks

Automatic Specimen Processing


Ionic Composition of Fixative Solution

Vehicle Osmolality

Effects of Added Substances

Methods for Adjusting the Osmolarity

Recommended Osmolality

Measurement of Osmolality


Buffer Types

Preparation of Buffers


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 of Glutaraldehyde




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


Reaction with Proteins

Reaction with Lipids

Reaction with Nucleic Acids

Reaction with Carbohydrates

Preparation of Paraformaldehyde


Reaction with Proteins

Reaction with Lipids

Precaution in the Handling of Acrolein

Acrolein Fixatives


Role of Aldehydes in Quantitative Electron Microscopy

Tissue Storage

4 Osmium Tetroxide


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


Preparations for and Precautions in the Handling of Osmium


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


Fixation Procedures

Permanganate Fixatives

6 Miscellaneous Fixatives

Ruthenium Tetroxide




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


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


10 Changes in Specimen Volume


11 Postmortem Changes


12 Plant Specimens


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


Duration of Fixation

Specimen Shrinkage


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




Mode of Fixation and Incubation

Single-Cell Specimens

Subcellular Fractions


15 Fixation for Immunoelectron Microscopy


16 Use of Dimethylsulfoxide in Fixation

Effect on Enzymes

Effect on Ultrastructure

17 Criteria for Satisfactory Specimen Preservation


18 Artifacts

Transmission Electron Microscopy

Scanning Electron Microscopy

19 Specific Fixation Methods



Balanced Salt Solutions

Commonly Used Salts and Their Physicochemical Properties

Commonly Used Chemicals




No. of pages:
© Academic Press 1981
28th December 1981
Academic Press
eBook ISBN:

About the Editor

M 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, USA

Ratings and Reviews