Keywords:
Flow cytometry.
;
DNA -- Analysis.
;
Electronic books.
Description / Table of Contents:
Cytometry is characterization and measurement of cells and cellular constituents, most often used to immunophenotype cells - that is, to distinguish healthy cells from diseased cells. Flow Cytometry specifically is quite sensitive, allowing researchers to detect rare cell types and residual levels of disease, and as such has been the method of choice for important studies such as monitoring the blood of AIDS patients. For this reason, there is a great need for a practical, comprehensive manual that will be useful across a broad range of laboratories. This volume, as part of the Reliable Lab Solution Series, delivers such a tool, offering busy researchers across many disciplines a handy resource of all the best methods and protocols for Cytometry to use at the bench. * Highlights top downloaded and cited chapters, authored by pioneers in the field and enhanced with their tips, and pitfalls to avoid. * Loaded with detailed protocols developed and used by leaders in the field. *Refines, organizes and updates popular methods from one of our top selling series, Methods in Cell Biology.
Type of Medium:
Online Resource
Pages:
1 online resource (945 pages)
Edition:
1st ed.
ISBN:
9780123750464
Series Statement:
Issn Series
URL:
https://ebookcentral.proquest.com/lib/geomar/detail.action?docID=566687
DDC:
571.6
Language:
English
Note:
Front Cover -- Essential Cytometry Methods -- Copyright Page -- Contents -- Contributors -- Preface -- Part I: Fluorochromes/General Techniques -- Chapter 1: Principles of Confocal Microscopy -- I. Brief History of Microscope Development -- II. Development of Confocal Microscopy -- III. Image Formation in Confocal Microscopy -- IV. Useful Fluorescent Probes for Confocal Microscopy -- V. Applications of Confocal Microscopy -- VI. Conclusions -- References -- Chapter 2: Protein Labeling with Fluorescent Probes -- I. Update -- II. Introduction -- III. Labeling of Proteins with Organic Fluorescent Dyes -- IV. Labeling of Antibodies with Zenon Probes -- V. Labeling of Proteins with Phycobiliproteins -- VI. Conclusion -- References -- Chapter 3: Cytometry of Fluorescence Resonance Energy Transfer -- I. Introduction -- II. Theory of FRET -- III. Measuring FRET -- IV. Applications -- V. Perspectives -- VI. Conclusions -- Acknowledgments -- References -- Chapter 4: The Rainbow of Fluorescent Proteins -- I. Update -- II. The Fluorescent Proteins -- III. Flow Analysis Using Fluorescent Proteins -- IV. Flow Sorting Using Fluorescent Proteins -- V. Conclusion -- Acknowledgments -- References -- Chapter 5: Labeling Cellular Targets with Semiconductor Quantum Dot Conjugates -- I. Introduction -- II. Selection of QDs and their Conjugates -- III. Labeling of Fixed Cells for Fluorescence Microscopy -- IV. Measurements in Living Cells Using QD Conjugates -- V. Conclusion -- Acknowledgment -- References -- Chapter 6: High-Gradient Magnetic Cell Sorting -- I. Update -- II. Introduction -- III. Application -- IV. Materials -- V. MACS: Staining and Sorting -- VI. Critical Aspects of the Procedure -- VII. Controls -- VIII. Instruments -- IX. Results -- Acknowledgment -- References -- Chapter 7: Multiplexed Microsphere Assays (MMAs) for Protein and DNA Binding Reactions.
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I. Update -- II. Introduction -- III. Technology and Instrumentation -- IV. Methods -- V. Results -- VI. Software -- VII. Critical Aspects of the Methodology -- VIII. Comparison with Other Methods -- IX. Future Directions -- Acknowledgment -- References -- Chapter 8: Biohazard Sorting -- I. Recent Developments -- II. Introduction -- III. Critical Aspects of the Procedure -- IV. Applications and Future Directions -- Acknowledgment -- References -- Chapter 9: Guidelines for the Presentation of Flow Cytometric Data -- I. Introduction -- II. General Principles of Graphical Presentation -- III. Statistics -- IV. Subset Analysis -- V. Conclusion -- References -- Part II: Cellular DNA Content Analysis -- Chapter 10: Detergent and Proteolytic Enzyme-Based Techniques for Nuclear Isolation and DNA Content Analysis -- I. Introduction -- II. Basic Principles of the Methods -- III. Applications -- IV. Materials -- V. Methods -- VI. Results -- Acknowledgments -- References -- Chapter 11: DNA Content and Cell Cycle Analysis by the Propidium Iodide-Hypotonic Citrate Method -- I. Introduction -- II. Application -- III. Materials -- IV. Staining Procedure -- V. Caution -- VI. Instrument Setup -- VII. Comments -- VIII. Results -- References -- Chapter 12: DNA Analysis from Paraffin-Embedded Blocks -- I. Introduction -- II. Application -- III. Methods -- IV. Critical Aspects of Technique -- V. Alternative Methods for Sample Preparation -- Acknowledgment -- References -- Chapter 13: Flow Cytometry and Sorting of Plant Protoplasts and Cells -- I. Update -- II. Introduction -- III. Application -- IV. Materials -- V. Procedures -- VI. Critical Aspects of the Procedures -- VII. Controls and Standards -- VIII. Instruments -- IX. Results -- Acknowledgments -- References -- Chapter 14: DNA Content Histogram and Cell-Cycle Analysis -- I. Introduction.
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II. DNA Content Histogram Basic Principles -- III. Cell-Cycle Analysis of DNA Content Histograms -- IV. Critical Aspects of DNA Content and Cell-Cycle Analysis -- V. Interpretation of DNA Content and Cell-Cycle Histograms -- VI. Conclusion -- References -- Chapter 15: Simultaneous Analysis of Cellular RNA and DNA Content -- I. Update -- II. Introduction -- III. Applications -- IV. Materials -- V Staining Procedures Employing AO -- VI. Staining RNA and DNA with PY and Hoechst 33342 -- VII. Critical Aspects of the Procedures -- VIII. Controls and Standards -- IX. Instruments -- X. Results -- XI Comparison of the Methods -- References -- Part III: Cell proliferation and the Death Assays -- Chapter 16: Immunochemical Quantitation of Bromodeoxyuridine: Application to Cell-Cycle Kinetics -- I. Update -- II. Introduction -- III. Applications -- IV. Materials -- V. Procedures -- VI. Critical Aspects of the Procedure -- VII. Controls and Standards -- VIII. Instruments -- IX. Results -- X. Summary -- References -- Chapter 17: Cell-Cycle Kinetics Estimated by Analysis of Bromodeoxyuridine Incorporation -- I. Introduction -- II. Applications -- III. Materials -- IV. Methods -- V. Critical Aspects of the Procedure -- References -- Chapter 18: Flow Cytometric Analysis of Cell Division History Using Dilution of Carboxyfluorescein Diacetate Succinimidyl Ester, a Stably . -- I. Introduction and Background -- II. Reagents and Solutions -- III. Preparation and Labeling of Cells -- IV. Gathering of Information Concurrent with Division -- V. Analysis of Data -- VI. Application of Carboxyfluorescein Diacetate Succinimidyl Ester to In Vitro Culture of Lymphocytes -- VII. Monitoring Lymphocyte Responses In Vivo -- VIII. Antigen Receptor Transgenic Models -- References.
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Chapter 19: Antibodies Against the Ki-67 Protein: Assessment of the Growth Fraction and Tools for Cell Cycle Analysis -- I. Introduction -- II. Application -- III. Materials and Methods -- IV. Critical Aspects -- V. Controls and Standards -- VI. Examples of Results -- References -- Chapter 20: Detection of DNA Damage in Individual Cells by Analysis of Histone H2AX Phosphorylation -- I. Update -- II. Introduction -- III. Considerations in the Use of gammaH2AX as a Measure ofDouble-Stranded Breaks -- IV. Methods of Analysis -- V. Typical Results -- VI. Possible Applications -- VII. Conclusion -- References -- Chapter 21: Assays of Cell Viability -- I. Update -- II. Introduction -- III. Changes in Light Scatter During Cell Death -- IV. Cell Sensitivity to Trypsin and DNase -- V. Fluorescein Diacetate (FDA) Hydrolysis and PI Exclusion -- VI. Rh123 Uptake and PI Exclusion -- VII. PI Exclusion Followed by Counterstaining with Hoechst 33342 -- VIII. Hoechst 33342 Active Uptake and PI Exclusion -- IX. Controlled Extraction of Low MW DNA from Apoptotic Cells -- X. Sensitivity of DNA In Situ to Denaturation -- XI. Detection of DNA Strand Breaks in Apoptotic Cells -- XII. A Selective Procedure for DNA Extraction from Apoptotic Cells Applicable to Gel Electrophoresis and Flow Cytometry... -- XIII. Comparison of the Methods: Confirmation of the Apoptotic Mode of Cell Death -- References -- Chapter 22: Difficulties and Pitfalls in Analysis of Apoptosis -- I. Update -- II. Introduction -- III. AI may not be Correlated with Incidence of Cell Death -- IV. Difficulties in Estimating Frequency of Apoptosis by Analysis of DNA Fragmentation -- V. The Lack of Evidence is not Evidence for the Lack of Apoptosis -- VI. Misclassification of Apoptotic Bodies or Nuclear Fragments as Single Apoptotic Cells -- VII. Apoptosis Versus Necrosis Versus ``Necrotic Stage´´ of Apoptosis.
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VIII. Selective Loss of Apoptotic Cells During Sample Preparation -- IX. Live Cells Engulfing Apoptotic Bodies Masquerade as Apoptotic Cells -- X The Problems with Commercial Kits and Reagents -- XI. Cell Morphology is still the Gold Standard for Identification of Apoptotic Cells -- XII. Laser Scanning Cytometry: Have your Cake and Eat it too -- References -- Part IV: Cell Surface Immunophenotyping -- Chapter 23: Cell Preparation for the Identification of Leukocytes -- I. Introduction -- II. Antibodies -- III. Tandem Fluorochromes -- IV. Cell Preparation and Staining Procedures -- V. Titering Antibodies -- VI. Solutions and Reagents -- References -- Chapter 24: Multicolor Immunophenotyping -- I. Update -- II. Introduction -- III. Methodology -- IV. Normal Immunophenotypic Patterns of Maturation -- V. Abnormal Immunophenotypic Patterns of Maturation -- References -- Chapter 25: Differential Diagnosis of T-Cell Lymphoproliferative Disorders by Flow Cytometry Immunophenotyping. Correlation with Morpholog. -- I. Introduction -- II. Materials -- III. Methods -- IV. Identification of Abnormal T-Cell Population by Flow Cytometry -- V. Precursor T-Lymphoblastic Lymphoma/Leukemia (T-ALL) -- VI. Peripheral (Mature/Postthymic) Lymphoma Versus T-ALL -- VII. Thymocytes from Thymic Hyperplasia/Thymoma Versus T-ALL -- VIII. Mature T-Cell Lymphoproliferative Disorders -- IX. Conclusions -- References -- Chapter 26: B Cell Immunophenotyping -- I. Update -- II. Introduction -- III. Immunophenotyping of B Cell Developmental Stages in the Bone Marrow -- IV. Peripheral B Cell Populations -- V. Summary -- VI. Antigen-Induced B Cell Subsets -- VII. Antigen-Specific B Cells -- References -- Part V: Cytogenetics/Chromatin Structure -- Chapter 27: Telomere Length Measurements Using Fluorescence In Situ Hybridization and Flow Cytometry -- I. Introduction -- II. Background.
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III. Methods.
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