Advanced Techniques in Biological Electron Microscopy II: Specific Ultrastructural Probes

The use of the term "advanced" in the title of this book is somewhat ar­ bitrary and very much relative with respect to time. Many techniques which were considered at the "cutting edge" of ultrastructural methodology just a few years ago are now rou tin ely used in numerous laboratories. One could cite freeze-fracture, cryothin sectioning, or indeed most of the field of scan­ ning electron microscopy as concrete examples. Thus the use of the term "ad­ vanced techniques" must be interpreted with regard to the present state of the art, and is useful only in informing the potential reader that this volume is not a primer to be used as an initial introduction into basic biological elec­ tron microscopy. Many excellent volumes have filled that niche in the past few years, and it is not intended that this modest book be a complete com­ pendium of the field. Furthermore, any limited selection of papers on advanc­ ed techniques necessarily reflects the preferences and arbitrary whims of the editor, thereby excluding many equally important procedures which the knowledgeable reader will readily identify. The first volume of this series appeared approximately five years ago and illustrated techniques which were thought to represent advanced and yet ba­ sically morphological methods for gaining increased ultrastructural informa­ tion from biological specimens. The present volume, on the other hand, stresses techniques which provide specific physicochemical data on the speci­ mens in addition to the structural information.

Table of Contents:
Ultrastructural Localization of Lectin Receptors.- A. Introduction.- B. Purification of Lectins.- C. Purification of Markers.- D. Synthesis of Probes and Labeling Techniques.- References.- Antibody-labeling Techniques.- A. Rationale.- B. Antibody Labels.- C. Methods for Coupling Label to Antibody.- D. Iodination of Antibody.- E. Hemocyanin Label of Antibody.- F. Reaction of Antibody with Cells.- G. Clotting Procedure for Handling Single Cell Suspensions.- H. Radioautography.- I. Replica Techniques.- J. Conclusions.- References.- Cell Surface Labeling for the Scanning Electron Microscope.- A. Introduction.- B. Labeling Techniques for the SEM.- C. Interpretation of Cell Surface Labeling in the SEM.- D. Summary.- References.- Low-Temperature Biological Scanning Electron Microscopy.- A. Introduction.- B. Low-Temperature Solidification of Cell and Tissue Fluids.- C. Pre-treatment Before the Cooling Process.- D. Specimen Cooling.- E. Post-freezing Preparative Procedures.- F. Specimen Transfer.- G. Low-temperature Specimen Stages.- H. Specimen Examination.- I. Conclusions.- References.- Quantitative Electron Microscopy of Nucleic Acids.- A. Introduction.- B. Basic Protein Film Method.- C. Heteroduplex Molecules.- D. Measuring and Error Analysis.- E. Artifacts and Topology Considerations.- F. RNA and Transcription Complexes.- G. Tagging Methods.- F. General Comments and Problems.- H. Protein-free Spreading.- References.- Electron Microscopy of Specific Proteins: Three-Dimensional Mapping of Ribosomal Proteins Using Antibody Labels.- A. Introduction.- B. Techniques.- C Interpretation.- References.- Electron Microscopy and Electron Diffraction Studies on Hydrated Membranes.- A. Introduction.- B. Operation of Hydration Chamber in an Electron Microscope.- C. Electron Microscopy.- D. Electron Diffraction (ED).- E. Conclusions and Future Development.- References.