Histochemistry

1 An Overview of Biological Staining 1.1 What Information Does Biological Staining Provide? 1.2 How Do We Carry out Biological Staining? And Why Do We Do What We Do? 1.3 What is Present to Be Stained? Fixation and Its Alternatives 1.4 Why Does Anything Become Stained? An Account of Reagent-Tissue Affinities, and of Other Causes 1.5 Why Doesn't Everything Always Stain? 1.6 Why Does the Tissue Remain Stained after Its Removal from the Staining Solution? 1.7 How Do We Judge the Significance of Staining? Or Indeed, of Failure to Stain?2 The Preparation of Material to Be Stained3 Fixation and Other Loss-Limiting Procedures 3.1 Why We Need to Limit Losses: The Practical Options 3.2 Limitation of Losses without Chemical Modification of the Tissue 3.3 Fixatives: Substances Used to Chemically Modify Native Tissues 3.4 How is the Fixation of Tissues Achieved? 3.5 The Fixation of Substances Other Than Protein 3.6 Effects of Fixation on Biological Staining: Limitation of Losses of Tissue Materials 3.7 Effects of Fixation on Biological Staining: Increases In Staining and Changes in Staining Patterns Due to Chemical Modifications of the Tissues 3.8 Effects of Fixation on Biological Staining Due to Physical Modifications of the Tissues 3.9 Rates of Fixative Penetration4 Staining Methods Involving Dyeing 4.1 What are Dyes and What is Dyeing? Definitions, Names and Descriptive Terms 4.2 Why are Dyes Colored? The Color, Dichroism, Fluorescence and Metachromasia of Dyes 4.3 Why Do Dyes Stain Anything? The Affinity Concept and Some Thermodynamic Formalism 4.4 Various Affinity Models for Biological Staining Model 1: Solvent Non-Aqueous, Dye Non-Ionic, No Dye-Tissue Forces Model 2: Solvent Non-Aqueous, Dye Non-Ionic, But Dye-Tissue Forces Occur: Van Der Waals Attractions, Hydrogen Bonding, Mordanting Model 3: Aqueous Solvent, Dye and/or Tissue Non-Ionic, No Appreciable Dye-Tissue Attractions: Hydrophobic Bonding Model 4: Dye and Substrate Ionic, All Other Variables Taken as Trivial Model 5: Aqueous Solvent, Dye and Substrate Ionic, Dye-Dye Attractions Occur 4.5 Dye-Tissue Affinities in Numbers: Various Quantitative Measures 4.6 Sources of Affinity - A Practical Summary 4.7 Why Don't Dyes Stain Everything? 4.8 Preliminaries Concerning Substrate Density and the Color of Staining 4.9 A Dye May Prefer One Substrate to Another: Some Affinity Models 4.10 Some Substrates Stain or Destain Faster than Others 4.11 What are the Causes of Differential Staining Rates? 4.12 How is the Rate of Staining Affected by the Dye Used? 4.13 How Do the Various Tissue Substrates Influence Staining Rates? 4.14 How Does Substrate Geometry Influence Staining Rate? 4.15 How Do Solvent and Co-Solute Affect Staining Rates? 4.16 How Do Pre-staining Treatments of the Tissue Affect Staining Rates? 4.17 A Summary of the Causes of Differential Rates of Staining 4.18 Various Examples of Rate-Controlled Selective Staining Procedures 4.19 Rate-Controlled Selective Staining Involving "Colorless Dyes" 4.20 Everything May Stain, and Stain at the Same Rate - But in a Variety of Colors5 Reactive Staining Methods 5.1 Some Definitions, Generalizations and Cautions 5.2 The Influence of Reagent Uptake on the Selectivity of Staining 5.3 The Influence of Reactivity on Reagent Selectivity6 Staining Methods Involving Uptake of Metals 6.1 A General Model for Staining Methods Involving Uptake of Metal 6.2 How Does the Uptake of Metal Occur? 6.3 How is the Metal Visualized after Uptake? 6.4 Why Should Such Methods Display Selectivity? 6.5 Why are Many Metal Impregnation Methods So Fickle?7 Enzyme Histochemistry 7.1 Essential Problems and Characteristics of Enzyme Histochemistry 7.2 Ultrastructural Localization of Enzymes versus Preservation of Their Biochemical Integrity 7.3 Substrates and Incubation Media 7.4 Formation and Properties of the Initial Reaction Product 7.5 Characteristics of the Capture-Visualization Steps 7.6 Properties of the Final Reaction Product 7.7 How Do We Interpret the Results of a Staining Procedure? 7.8 Peculiarities of Electron-Microscopic Enzyme Histochemistry8 Immuno-Chemical Staining Using Labeled Antibody and Related Methods 8.1 Some Strategies of Staining 8.2 A Sketch of the Chemistry of Antibodies and Antigens, and of Their Interactions 8.3 Preparation of Antibodies and of Analogous Reagents 8.4 Nature and Preparation of the Label 8.5 Labeling Methods: Preparation of Labeled Antibodies and Related Materials 8.6 Nature of Visualizing Procedures 8.7 Specificity of the Staining 8.8 Some Factors Limiting Sensitivity 8.9 The Conflicting Problems of Preserving Antigen and Antigenicity (Or Antibody and Immunogenicity) and Cell Morphology9 Some Miscellaneous Biological Staining Methods 9.1 An Introduction 9.2 Dyeing with Reagents That are Not Dyes 9.3 A Trapping Method: The Gram Reaction 9.4 Vital Staining Procedures10 Peculiarities of Staining Plastic Sections 10.1 The State of the Art of Staining Plastic Sections 10.2 What is the Effect of the Fixative?10.3 Some Properties and Chemistry of Plastic Embedding Media, and of the Relevant Monomers 10.4 Penetration of Reagents into Plastic Sections: Some Observations and a Rationale 10.5 A General Approach to Staining Plastic Sections, with Some Applications and Complications 10.6 Some Practical Implications of the General Model11 The Interpretation and Analysis of Histochemical Observations 11.1 Checking the Meaning of Staining: An Introduction 11.2 The Use of Selective Extraction 11.3 The Use of Histochemical Blocking 11.4 The Use of Other Control Procedures 11.5 Does Staining Intensity Reflect Substrate Concentration? 11.6 Is the Stain Where the Substrate Was? The Localization Problem 11.7 Checking the Meaning of No-Staining: An Introduction 11.8 Have There Been Substrate Losses? 11.9 Is There Masking of Substrate? 11.10 Was There Staining, Which Has Been Lost? 11.11 The Inevitability, Value and Nature of Staining Theories 11.12 Uses and Abuses of Test Objects, Model Systems and Histochemical-Chemical Comparisons as Aids to Understanding Staining Results 11.13 The Baleful Influence of Reagent Impurities on the Interpretation of Staining Results 11.14 The Uses of Structure-Staining Correlations, Practical and Theoretical 11.15 A Summary of Some Practical Strategies12 Supplements 12.1 Nomenclature of Traditional Color Theory: Chromagen, Chromophore and Auxochrome 12.2 Π Bonds and Σ Bonds: A Glance at the Valency of Carbon 12.3 Relation of Affinity to Other Thermodynamic Quantities 12.4 The Nature and Characteristics of Van Der Waals Attractions 12.5 The Hydrogen Bond 12.6 The Coordinative Bond 12.7 Hydrophobic Bonding 12.8 Quantitative Estimation of Affinity: Use of Freundlich and Langmuir Isotherms 12.9 Methods of Determining Half Staining Times and Activation Energies for the Staining of Tissue Sections 12.10 Aggregation of Dyes in Solution: Some Physical Chemistry 12.11 The Precipitation of Sparingly Soluble ("Insoluble") Salts 12.12 Some Structural Characteristics of Immunoglobulins 12.13 Photofading: Some Physico-Chemical Background to the Fading of Stained Preparations Following Exposure to Visible or Ultra Violet RadiationReferencesSubject Index