Cell Biology
A Laboratory Handbook
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Cell Biology: A Laboratory Handbook, Volume 3 is a handbook on cell biology and covers topics ranging from transfer of macromolecules and small molecules to cloning of embryos, transgenics, and gene targeting. Cell-free extracts, permeabilized cell systems, and expression systems are also discussed, along with proteins. Comprised of 58 chapters, this volume begins with a detailed account of microinjection of RNA, DNA, and proteins into somatic cells, followed by an analysis of computer-automated capillary microinjection of macromolecules into living cells. The reader is then introduced to syringe loading as a method for inserting macromolecules into cells in suspension; electroporation of cells; and the use of liposomes in drug targeting. Subsequent chapters focus on the cloning of rabbit embryos by nuclear transplantation; gene targeting by homologous recombination in embryonic stem cells; production and isolation of recombinant viruses; and gel electrophoresis. This book will be of interest to geneticists and molecular biologists.
Table of Contents
Part 1: Tissue Culture and Associated Techniques
Section A: General Techniques
General Procedures for Tissue Culture
Development of Serum-Free Media and Methods for Optimization of Nutrient Composition
Testing Cell Cultures for Microbial and Viral Contaminants
Section B: Primary Cultures from Embryonic Tissues
Primary and Extended Culture of Embryonic Mouse Cells: Establishment of a Novel Cell Culture Model of Apoptosis and Neural Differentiation
Tissue Culture of Embryonic Stem Cells
Isolation and Culture of Germ Cells from the Mouse Embryo
Section C: Cultures of Specific Cell Types
Epithelial Cells
Cultivation of Human Epidermal Keratinocytes with a 3T3 Feeder Layer
Growth of Human Keratinocytes in Serum-Free Medium
Isolation of Hepatocytes
Isolation and Culture of Oval Cells from Carcinogen-Treated Rats
In Vitro Culture of Mouse Fetal Choroid Plexus Epithelial Cells
Isolation and Culture of Type II Pulmonary Epithelial Cells
Mesenchymal Cells
Maintenance of Human Diploid Fibroblast-Like Cells in Culture
Isolation of Osteoclasts and Osteoclast Plasma Membranes
Culturing of Human Umbilical Vein and Dermal Microvascular Endothelial Cells
Neuroectodermal Cells
Isolation and Proliferation of Adult Mammalian Central Nervous System Stem Cells
Hemopoietic Cells
Clonal Cultures in Vitro for Hemopoietic Cells Using Semisolid Agar Medium
Gonads
Properties of Isolated Sertoli Cells
Culture of Ovarian Granulosa Cells: Calcium Imaging at the Single-Cell Level
Section D: Cell Separation Techniques
Isolation of Peripheral Blood Mononuclear Cells and Identification of Human Lymphocyte Subpopulations by Multiparameter Flow Cytometry
Purification of Functionally Active Epidermal Langerhans Cells Using Immunomagnetic Beads
Section E: Model Systems to Study Differentiation
Nonterminal and Terminal Adipocyte Differentiation of Murine 3T3 T Mesenchymal Stem Cells
Cell Systems for ex Vivo Studies of Myogenesis: A Protocol for the Isolation of Stable Muscle Cell Populations from Newborn to Adult Mice
Induction of Cell Differentiation in Human HL-60 Promyelocyte Leukemia Cells: Quantitation of a Myeloid Specific Antigen, MRP-8/MRP-14 Protein Complex
Differentiation of Murine Erythroleukemia Cells (Friend Cells)
Cultured PC 12 Cells: A Model for Neuronal Function and Differentiation
Growing Madin-Darby Canine Kidney Cells for Studying Epithelial Cell Biology
In Vitro Studies of Epithelium-to-Mesenchyme Transitions
Section F: Immortalization of Cells
Inducible Immortalization of Cells from Transgenic Mice Expressing Simian Virus 40 under lac Operon Control
Immortalization of Rat Ventral Prostate Epithelial Cells Using Simian Virus 40 T Antigen
Section G: Cell Cycle Analysis
Cell Cycle Analysis by Flow Cytometry
Preparation of Synchronous Populations of Mammalian Cells in Specific Phases of the Cell Cycle by Centrifugal Elutriation
Synchronization of Normal Diploid and Transformed Mammalian Cells
Synchronization of Transformed Human Amnion Cells by Mitotic Detachment
Stimulation of DNA Synthesis in Quiescent 3T3 Cells
Section H: Cytotoxic Assays
Quantitative Determination of Compound Cytotoxicity in Proliferating Cells: Monitoring DNA Synthesis by [3H]Thymidine Incorporation
Section I: Senescence, Programmed Cell Death, and Others
Serial Propagation of Human Fibroblasts for the Study of Aging at the Cellular Level
Morphological Criteria for Identifying Apoptosis
Use of the Terminal Transferase DNA Labeling Reaction for the Biochemical and in Situ Analysis of Apoptosis
Growth and Induction of Metastasis of Mammary Epithelial Cells
Measurement of Cell-Cell and Cell-Extracellular Matrix Interactions: A Quantitative Cell Attachment Assay
Section J: Electrophysiological Methods
Patch-Clamp Recording
Section K: Histocultures
Three-Dimensional Sponge-Gel Matrix Histoculture: Methods and Applications
Section L: Other Cell Types
Anthropoda
Primary Culture of Drosophila Embryo Cells
Caenorhabditis elegans
Laboratory Cultivation of Caenorhabditis elegans and Other Free-Living Nematodes
Protozoa
Cultivation of Tetrahymena Cells
Acanthamoeba castellanii: A Model System for Correlative Biochemical and Cell Biological Studies
Fungi
Cell Biological, Molecular Genetic, and Biochemical Methods to Examine Dictyostelium
Large-Scale Culture of Physarum: A Simple Method for Growing Several Hundred Grams of Plasmodia
Plants
Induction of Regeneration-Competent Monocot Callus
Isolation, Culture, and Plant Regeneration from Protoplasts
Part 2: Viruses
Propagation and Purification of Polyoma and Simian Virus 40
Construction and Propagation of Human Adenovirus Vectors
Tissue Culture Techniques for the Study of Human Papilloma Viruses
Growth and Purification of Murine Leukemia Virus
Part 3: Organelles, Cellular Structures, Macromolecules, and Functional Assays
Purification of Rat Liver Golgi Stacks
Preparation and Purification of Post-Golgi Transport Vesicles from Perforated Madin-Darby Canine Kidney Cells
Purification of Clathrin-Coated Vesicles from Bovine Brain, Liver, and Adrenal Gland
Functional Identification of Membranes Derived from the Rough Endoplasmic Reticulum of Yeast
Isolation of Yeast Mitochondria and Study of Mitochondrial Protein Translation
Inclusion of Proteins into Isolated Mitochondrial Outer Membrane Vesicles
Isolation of Peroxisomes
Purification of Secretory Granules from PC 12 Cells
Preparation of Synaptic Vesicles from Mammalian Brain
Purification and Reconstitution of the Ca2+ -ATPase of Red Blood Cells
Isolation of Focal Adhesions from Cultured Cells
Isolation of Laminins from Tumor Sources and from Normal Tissues
Isolation of Centrosomes from Cultured Animal Cells
Preparation of Yeast Spindle Pole Bodies
Preparation of Nuclei and Nuclear Envelopes, Identification of an Integral Membrane Protein Unique to the Nuclear Envelope
Preparation of Cytoplasts and Karyoplasts from HeLa Cell Monolayers
Isolation and Visualization of the Nuclear Matrix, the Nonchromatin Structure of the Nucleus
Preparation of U Small Nuclear Ribonucleoprotein Particles
Rapid Preparation of hnRNP Core Proteins and Stepwise Assembly of hnRNP Particles in Vitro
Preparation of Ribosomes and Ribosomal Proteins from Cultured Cells
Preparation of Proteasomes
Small-Scale Preparation of Nuclear Extracts from Mammalian Cells
Purification of DNA Using Guanidine Thiocyanate and Isobutyl Alcohol Fractionation
Single-Step Method of Total RNA Isolation by Acid Guanidine-Phenol Extraction
Part 4: Microscopy Techniques
Section A: Light Microscopy
Phase-Contrast, Nomarski (Differential-Interference) Contrast, and Dark- Field Microscopy: Black and White and Color Photomicrography
Reflection Interference Microscopy
Using Interference Microscopy to Study Cell Behavior
Section B: Fluorescence Microscopy
Fluorescence Imaging in Living Cells
Practical Laser-Scanning Confocal Light Microscopy: Obtaining Optimal Performance from Your Instrument
Caged Fluorescent Probes for Monitoring Cytoskeleton Dynamics
Section C: Video Microscopy
Video-Enhanced Contrast Microscopy
Section D: Confocal Microscopy
Confocal Microscopy of Polarized MDCK Epithelial Cells
Real-Time Confocal Microscopy and Cell Biology
Section E: Electron Microscopy
Fixation of Cells and Tissues for Transmission Electron Microscopy
Embedding of Cells and Tissues for Ultrastructural and Immunocytochemical Analysis
Negative Staining
Whole-Mount Electron Microscopy of the Cytoskeleton: Negative Staining Methods
Glycerol Spraying/Low-Angle Rotary Metal Shadowing
Rapid Freezing of Biological Specimens for Freeze-Fracture and Deep Etching
Freeze Fracture and Freeze Etching
Use of Ultrathin Cryo- and Plastic Sections for Immunoelectron Microscopy
Cryo-Transmisson Electron Microscopy of Thin Vitrified Sections
Preparation Methods for Quantitative X-Ray Microanalysis of intracellular Elements in Ultrathin Sections for Transmission Electron Microscopy: The Freeze-Dry, Resin-Embed Route
High-Resolution Scanning Electron Microscopy in Cell Biology
Part 5: Microdissection Techniques
Microdissection-Based Techniques for the Determination of Cell Proliferation in Gastrointestinal Epithelium: Application to Animal and Human Studies
Micromanipulation of Chromosomes Using Laser Microsurgery (Optical Scissors) and Laser-Induced Optical Forces [Optical Tweezers)
Microdissection of Chromosomes and Microcloning
Part 6: Histochemistry
Mayer's Hematoxylin-Eosin: An Example of a Common Histological Staining Method
Selected Enzyme Staining in Histochemistry
Part 7: Antibodies
Section A: Production of Antibodies
Production of Polyclonal Antibodies in Rabbits
Production of Mouse Monoclonal Antibodies
Production of Human Monoclonal Antibodies via Fusion of Epstein-Barr Virus-Transformed Lymphocytes with Heteromyeloma
Rapid Production of Antibodies in Chicken and Isolation from Eggs
Section B: Purification and Labeling of Immunoglobulins
Purification of Immunoglobulins
Conjugation of Fluorescent Dyes to Antibodies
Section C: Antibody Specificity
Determination of Antibody Specificity by Western Blotting and Immunoprecipitation
Western Blotting and Ligand Blotting Using Enhanced Chemiluminescence and Radioiodine Detection
Enzyme-Linked Immunosorbent Assay
A Simple Solid-Phase Mutual Inhibition Assay Using Biotinylated Antigen for Analyzing the Epitope Specificities of Monoclonal Antibodies
DNA Immunoprecipitation: Application to Characterization of Target Sequences for a Human Centromere DNA-binding Protein (CENP-B)
Part 8: Immunocytochemistry And Vital Staining of Cells
Immunofluorescence Microscopy of Cultured Cells
Immunofluorescence Microscopy of the Cytoskeleton: Double and Triple Immunofluorescence
Immunocytochemistry of Frozen and of Paraffin Tissue Sections
Fluorescent Labeling of Nascent RNA in the Cell Nucleus Using 5-Bromouridine 5'-Triphosphate
Labeling of Endocytic Vesicles Using Fluorescent Probes for Fluid-Phase Endocytosis
Labeling of the Endoplasmic Reticulum with DiO6 (3)
Use of Fluorescent Analogs of Ceramide to Study the Golgi Apparatus of Animal Cells
Vital Staining of Mitochondria with Rhodamine 123 and of Acidic Organelles with Acridine Orange
Part 9: Intracellular Measurements
Measuring Membrane Potential in Single Cells with Confocal Microscopy
Measurement of Cytosolic pH in Single Cells by Dual-Excitation Fluorescence Spectrometry: Simultaneous Visualization Using Hoffman Modulation Contrast Optics
Part 10: Cytogenetics and in Situ Hybridization
Basic Cytogenetic Techniques: Culturing, Slide Making, and G-Banding
Production of Viable Hybrids Between Adherent Cells
Microcell-Mediated Chromosome Transfer: Selective Transfer and Retention of Single Human Chromosomes into Recipient Cells of Choice
Microcell Transfer of Chromosomes from Mitotic Cells
Chromosome Painting Using Degenerate Oligonucleotide-Primed Polymerase Chain Reaction-Amplified, Flow-Sorted Human Chromosomes
Fluorescence in Situ Hybridization of Human and Mouse DNA Probes to Determine the Chromosomal Contents of Cell Lines and Tumors
in Situ Hybridization Applicable to Abundantly Expressed mRNA Species
In Situ Hybridization of Frozen Sections Using 3 5S-Riboprobes
In Situ Detection of Human Papillomavirus DNA After Polymerase Chain Reaction Amplification
Accurate Quantitation of mRNA Species by Polymerase Chain Reaction and Solid-Phase Minisequencing
Product details
- No. of pages: 566
- Language: English
- Copyright: © Academic Press 1994
- Published: January 1, 1994
- Imprint: Academic Press
- eBook ISBN: 9781483282459