Practical Methods in Electron Microscopy

In this latest volume in the series detailed practical guidance is given for the preparation of ultrathin sections for electron microscopy and of thicker sections for light microscopy from blocks of resin-embedded material. Techniques for the sectioning of the newer acrylic resins and the use of diamond knives are fully described, and methods for overcoming sectioning problems are discussed in detail.

Table of Contents:
Editor's preface. Authors' preface. Acknowledgements. Chapter 1: Introduction. 1.1 The scope of ultramicrotomy. 1.2 The theory of sectioning. 1.2.1 The knife. 1.2.2 Section distortion. 1.2.3 Production of heat during sectioning. 1.2.4 Interactions between the knife and the specimen block. 1.2.5 Sectioning in practice. 1.3 How thick should a section be? 1.4 Serial sections. Chapter 2: Microtomes and ultramicrotomes. 2.1 The development of sectioning instruments. 2.2 Principles of ultramicrotome design. 2.2.1 The advance mechanism. 2.2.2 The cutting operation. 2.2.3 Other features of the ultramicrotome. 2.3 Ultramicrotomes. 2.3.1 Cambridge Huxley ultramicrotome. 2.3.2 Sorvall ultramicrotomes. 2.3.3 LKB Ultrotome ultramicrotomes. 2.3.3a LKB Ultrotome V, LKB Nova and Reichert-Jung Super Nova. 2.3.3b LKB Ultrotome IV. 2.3.4 Reichert-Jung ultramicrotomes. 2.3.5 Choosing an ultramicrotome. 2.4 Supports and tables for ultramicrotomes. 2.5 Microtomes. 2.5.1 Sorvall JB-4 microtome. 2.5.2 Reichert-Jung microtomes. 2.5.3 Reichert-Jung Polycut microtome. 2.5.4 LKB Historange microtome. 2.5.5 Bright microtome. 2.5.6 Anglia Scientific microtome. Chapter 3: Knives. 3.1 The requirements of a knife for ultramicrotomy. 3.2 Glass knives. 3.2.1 Glass for knife making. 3.2.2 Glass strips. 3.3 Triangular glass knives for ultrathin sectioning. 3.3.1 The positioning of the score. 3.3.2 The quality of the knife edge. 3.3.3 Methods of making glass knives. 3.3.4 The knife angle. 3.3.5 Knife edge evaluation. 3.3.6 Tungsten-coated glass knives. 3.4 Knife-making instruments for triangular glass knives. 3.4.1 LKB KnifeMakers. 3.4.1a LKB 7800 KnifeMaker. 3.4.1b LKB 2178 KnifeMaker II. 3.4.2 Sunkay Messer knife maker. 3.4.3 Taab-Pyper glass knife maker. 3.4.4 Bio-Rad (Sorvall) glass knife maker. 3.4.5 Nisshin EM knife makers. 3.5 Histo-knives or Ralf knives. 3.6 Histo knife-making instruments. 3.6.1 LKB Histo KnifeMaker. 3.6.2 Taab Histo knife maker (or Reichert-Jung Histo knife maker). 3.6.3 Pelco 'Longknife' maker. 3.7 Adapters for histo-knives. 3.8 Diamond knives. 3.8.1 Use of a diamond knife. 3.8.2 Cleaning a diamond knife. 3.9 Troughs for glass knives. 3.10 Storage of knives. Chapter 4: Semithin and thick sections. 4.1 Uses of semithin and thick sections. 4.1.1 Semithin sections as an adjunct to electron microscopy. 4.1.2 Sections for high-voltage electron microscopy. 4.1.3 Survey sections. 4.1.3a Re-embedding methods for survey sections mounted on glass slides. 4.1.3b Mounting survey sections on dummy blocks. 4.1.3c Embedding survey sections in epoxy resin slides. 4.1.4 Sections for immunocytochemistry and autoradiography. 4.1.5 Sections for three-dimensional analysis. 4.2 The choice of embedding medium for semithin and thick sections. 4.3 The choice of section thickness. 4.4 Sectioning and staining procedures. 4.4.1 Dry sectioning of hydropholic blocks. 4.4.2 Sectioning of hydrophobic blocks. 4.4.3 Serial semithin and thick sections.