附註:Includes bibliographical references and index.
Intro; Contents; List of Contributors; Abbreviations; 1. Cell culture; 1. Introduction; Sources of cell cultures; Suppliers of cell cultures; Types of cell culture; 2. Culture media; Basal media; Serum; Serum-free media; Antibiotics; 3. Good practice in cell culture; 4. Principles of cell banking; 5. Subculture of cells; 6. Quantification of cell cultures; 7. Cryopreservation and storage of cell lines; Storage of cryopreserved cell stocks; Resuscitation of cryopreserved cell stocks; 8. Quality control testing of cell lines; Preparation of cell cultures prior to testing; Mycoplasma
Bacteria and fungiViruses; Isoenzyme analysis; Karyotyping; DNA fingerprinting; 9. Transportation; Shipment of growing cultures; Shipment of frozen materials on dry ice (solid carbon dioxide); References; 2. Virus isolation; 1. Why culture viruses?; 2. Viruses not amenable to cultivation; 3. Treatment of clinical specimens for virus isolation; 4. Identification of virus replication; 5. Passaging of viruses; 6. Confirmation of virus isolation; 7. Adaptations of cell culture; Haemadsorption test; Detection of early antigen fluorescent foci (DEAFF test); 8. Conclusions; References
3. Concentration and purification of viruses1. The need for virus concentration and purification; Practical considerations; 2. Virus concentration: general principles; 3. Purification of virus: identification of virus-containing fractions; 4. Purification by ultracentrifugation; Types of ultracentrifugation gradient; Forming gradients; 5. Assessment of purity; References; 4. Assays for virus infection; 1. Introduction; 2. Identification of viruses isolated in cell culture; Cytopathic effects; Virus infectivity assays; Immunofluorescence; Electron microscopy
Other methods of identifying viruses growing in cell cultureDetection of virus pre-cytopathic effects; Detection of viruses not producing a cytopathic effect; 3. Virus antigen detection; Immunofluorescence; Particle agglutination; Antigen-capture ELISA; 4. Virus genome detection; Nucleic acid extraction; Genome detection assays; Quantitative nucleic acid amplification techniques; 5. Detection of specific anti-viral antibodies; Virus-specific IgM detection; Detection of virus-specific IgG; References; 5. Electron microscopy of viruses; 1. Introduction
2. Virus structure and strategies for virus detection3. Specimen preparation; Centrifugation; Ultracentrifugation; Ultrafiltration; Precipitation; Bioaffinity techniques; 4. EM grids and support films; 5. Negative staining; Negative stains: properties; Negative staining procedures; Quantitative aspects: measuring and counting in TEM; 6. Embedding and thin sectioning; Fixation and staining; Embedding; Thin sectioning; Post-staining; 7. Immuno-electron microscopy (IEM); Antibodies: properties and usage; Immuno-negative staining; Immuno-labelling of thin sections
摘要:Viruses have properties that are distinct from other living organisms and so require different methods to culture them. This volume provides a broad treatment of the principles and practice of virus culture and will be of interest to all those involved in virus culture including academic, industrial, and clinical research groups.
