Histological Techniques For Electron Microscopy

Histological Techniques for Electron Microscopy, Second Edition, offers a practical guide for those who would study cells or tissues with an electron microscope. The book contains 11 chapters and begins with a discussion of the organization and management of an electron microscope laboratory. This is followed by separate chapters on tissue preservation; fixatives and their variations; methacrylate embedding and cross-linked plastics, microtomes and microtomy; and section mounting. Subsequent chapters deal with the techniques of "staining" for electron microscopy, photography; mounting, shadowing, and replication; and negative staining.

Electron Microscopy Methods and Protocols is designed for the established researcher as a manual for extending knowledge of the field. It is also for the newcomer who wishes to move into the field. A wide range of applications for the examination of cells, tissues, biological macromolecules, molecular structures, and their interactions are discussed. We have tried to gather together methods that we consider to be those most generally appli- ble to current research in both cell and molecular biology. Each chapter c- tains a set of related practical protocols with examples provided by experts who have first-hand knowledge of the techniques they describe. The individual chapters are grouped according to similarities in their specimen preparation and methodology. Methods are presented in detail, in a step-by-step fashion, using reproducible protocols the authors have personally checked. During the last decade, the scientific literature describing the use of colloidal gold as an immunocytochemical marker has increased at an ex- nential rate, and this trend is expected to continue. We have included a large number of variations on the immunogold labeling technique. In both the ne- tive staining and cryo chapters, authors emphasize the “immunological app- cations” in order to correlate as fully as possible with the emphasis on immunogold labeling in the other chapters. Electron Microscopy Methods and Protocols commences with the routine preparation of biological material for classical transmission electron microscopy involving tissue fixation, embedding, and sectioning (Chap. 1).

Freeze-substitution and freeze-drying; Freezing technology; The freeze-etching technique; Freeze-etching methodology; Intepretation of freeze-etching images; Negative staining; Equipment requirements; Negative staining; Summary comments on negative staining procedures; Photography of negatively stained specimens; Particle dimensions and molecular weights; Shadow casting and replication; High resolution and shadowing; Autoradiography; Conditions for quantitation; Analysis of autoradrograms; Technical considerations.