Gareth Griffiths

Fine Structure Immunocytochemistry

Mitarbeit: Burke, B.; Lucocq, J.

• Broschiertes Buch

Produktbeschreibung

Electron microscopy in the biological sciences can be divided into two disciplines. The first, concerned with high resolution detail of particles or periodic structures, is mostly based on sound theoretical principles of physics. The second, by far the larger discipline, is interested in the information obtainable from thin sections. The theoretical back ground to those groups of techniques for preparing and looking at thin sections is often inexact and "loose", for want of a better word. What should be chemistry is often closer to alchemy. This kind of electron microscopy is often enshrined with mystical recipes, handed down from generation to generation. Admittedly, many of the processes involved, such as those required to embed tissue in epoxy resins, involve multiple interconnected steps, which make it difficult to follow the details of anyone of these steps. If all these steps are shrouded in some mystery, however, can one really trust the final image that emerges on the EM screen? When we present the data in some semi quantitative form is there really no better way to do it than to categorize the parameters with ++, +/-, etc? What happens when one labels the sections with antibodies? Does the whole business necess arily need to be more of an "art" than a "science"? Upon reflecting on these problems in 1981, I had the impression that many of the multi-authored textbooks that existed then (and that have appeared since) tended to exacerbate or at least perpetuate this

Produktdetails

• Verlag: Springer / Springer Berlin Heidelberg / Springer, Berlin
• Artikelnr. des Verlages: 978-3-642-77097-5
• Softcover reprint of the original 1st ed. 1993
• Seitenzahl: 484
• Erscheinungstermin: 6. Dezember 2011
• Englisch
• Abmessung: 235mm x 155mm x 27mm
• Gewicht: 726g
• ISBN-13: 9783642770975
• ISBN-10: 3642770975
• Artikelnr.: 36121962

Inhaltsangabe

1 Introduction to Immunocytochemistry and Historical Background.- 1.1 Approaches to Immunocytochemistry.- 1.2 Criteria for an Electron-Microscopic Immunocytochemical Technique.- 1.3 Problems with the Pre-Embedding Techniques.- References.- 2 Fine Structure Preservation.- 2.1 Introduction.- 2.2 Fine-Structure Evaluation in Practice.- 2.3 Summary.- References.- 3 Fixation for Fine Structure Preservation and Immunocytochemistry.- 3.1 Fine Structure Preservation.- 3.2 Factors Affecting the Quality of Fixation for Fine Structure Preservation.- 3.3 Fixation Artefacts.- 3.4 Effect of Fixatives on Enzyme Activity.- 3.5 Fixation for Immunocytochemistry.- 3.6 A Concluding Remark.- References.- 4 Embedding Media for Section Immunocytochemistry.- 4.1 Resin-Free Sections - Temporary Embedement.- 4.2 Permanent Embedding Media.- 4.3 Freeze Substitution.- References.- Chapters 5 Cryo and Replica Techniques for Immunolabelling.- 5.1 Cryo Sectioning Techniques.- 5.2 Freeze-Fracture and Replica Labelling Methods.- 5.3 Other Replica Techniques for Labelling.- References.- 6 Elementary Immunology.- 6.1 Basic Immunology.- 6.2 The Structure of Antibodies.- 6.3 The Biological Functions of Immunoglobulins.- 6.4. The Nature of Antigenicity.- 6.5 Practical Aspects of Immunology.- 6.6 Affinity Purification.- 6.7 Characterization of Antibodies.- 6.8 Precipitin Techniques.- 6.9 Immunoblotting.- 6.10 Immunoprecipitation.- 6.11 Immunoassay.- References.- 7 Labelling Reactions for Immunocytochemistry.- 7.1 Historical Perspectives.- 7.2 Labelling in Practice.- 7.3 Approaches for Single Labelling.- 7.4 Approaches for Double Labelling.- References.- 8 Particulate Markers for Immunoelectron Microscopy.- 8.1 Colloidal Gold.- Appendix. Silver Enhancement Procedure.- References.- 9 Non-ImmunologicalHigh-Affinity Interactions Used for Labelling.- 9.1 Lectins.- 9.2 Protein A.- 9.3 Protein G.- 9.4 Avidin-Biotin Interactions.- 9.5 Miscellaneous Labelling Approaches Relying on Non-Immunological High Affinity Interactions.- 9.6 In-Situ Hybridization.- References.- 10 Preembedding Immuno-Labelling.- 10.1 Permeabilization.- 10.2 Markers for Preembedding Labelling.- 10.3 Preembedding Studies of the Nucleus.- 10.4 Preembedding Labelling for Studies of the Nervous System.- 10.5 The Use of Dinitrophenol IgG Conjugates.- 10.6 Agarose Gel Method for Preembedding.- References.- 11 Quantitative Aspects of Immunocytochemistry.- 11.1 General Comments.- 11.2 Basic Stereology.- 11.3 Estimation of Volume Density and Surface Density in Practice.- 11.4 Estimation of Volume of Reference Space.- 11.5 Stereological Sampling in Practice.- 11.6 Quantitation in Immunocytochemistry.- 11.7 Approaches for Quantitation Ignoring Labelling Efficiency.- 11.8 Absolute Quantitation Making Assumptions About Labelling Efficiency.- 11.9 Negation of the Labelling Efficiency Factor: The Matrix Gel Method for Quantitating Souble Antigens Which Are Available in Pure Form.- 11.10 Quantitative Model System Developed for Small Molecular Weight Neurotransmitters.- 11.11 Sensitivity of Labelling: Limits of Detection.- 11.12 Steric Hindrance.- 11.13 Amplification of Gold Labelling and Quantitation.- 11.14 Signal-to-Noise Ratio and the Concentration of Antigen: Those Few Gold Particles!.- References.- 12 An Overview of Techniques for Labelling at the EM Level.

Rezensionen

"a wealth of solid, well-referenced information...The more experienced user will find this is an essential reference text" Australian Journal of Medical Science