Hybridization Techniques for Electron MicroscopyCRC Press, 1993 M06 9 - 368 pages Hybridization Techniques for Electron Microscopy examines the use of in situ hybridization techniques, including an overview of current perspectives and future developments. The book features in situ methods for fluorescence probes and confocal scanning microscopes. Three in situ hybridization methods for electron microscopes are analyzed: the non-embedded tissue method using ultrathin frozen sections, pre-embedded method, and post-embedded method using material embedded in hydrophilic resin. Positive and negative features are discussed, and clear instructions regarding implementation of techniques are provided. Particular aspects of the techniques are examined in detail, such as preparation of tissue, pretreatment, hybridization procedures, revelation (autoradiography and immunocytology) and checking procedures, in addition to the illustration, interpretation, and discussion of methods and results. The main applications described include virus detection, chromosomal gene mapping, detection of ribosomic nucleic acid, and detection of messenger RNA in animals and plants. Hybridization Techniques for Electron Microscopy is an excellent reference for cytologists, cell biologists, histochemists, cytochemists, molecular endocrinologists, and neuroendocrinologists. |
Contents
Introduction | 1 |
G Morel | 10 |
Chapter 2 | 45 |
Chapter 3 | 62 |
Chapter 4 | 112 |
VI | 120 |
In situ Hybridization on Semithin Sections 99 | 126 |
Acknowledgments | 135 |
Chapter 5 | 139 |
Common terms and phrases
anti-biotin antibody antigen autoradiography biotin biotinylated biotinylated probes cDNA cellular chromosome colloidal gold colloidal gold particles conjugated Cytochem cytoplasm denaturation described in Protocol detection digoxigenin diluted distilled water DNA probes DNase double-stranded electron microscope embedding emulsion enzymatic enzyme ethanol Figure fixation formamide gene gentle agitation glutaraldehyde grids on buffer growth hormone Histochem hybridization buffer immunocytological Incubate the grids labeled probe Lowicryl K4M method microscope level molecules Morel mRNA nick translation nonradioactive nonspecific nucleic acid nucleotides nucleus oligonucleotide probe Ovalbumine paraformaldehyde phosphate buffer pituitary plasmid polymerase polymerization post-embedding pre-embedding Prepare pretreatment procedure protein Protocols 9 Puvion-Dutilleul radioactive reaction reagents resin Rinse RNase room temperature semithin sections sequences serum silver grains single-stranded situ hybridization slides sodium somatotrophs specific staining streptavidin technique tissue Ultrastructural localization ultrathin frozen sections ultrathin sections unlabeled uranyl acetate vibratome viral DNA viral genomes viroplasms virus Wash the grids µg/ml