Note: Front Cover -- Methods in Cell Biology, Volume 41 -- Copyright Page -- Contents -- Contributors -- Preface to the Second Edition -- Preface to the First Edition -- Chapter 1. Dissociation of Intact Cells from Tumors and Normal Tissues -- I. Introduction -- II. Application -- III. Methodology: Mechanical Cell Dispersion -- IV. Results -- V. Conclusion -- References -- Chapter 2. Assays of Cell Viability: Discrimination of Cells Dying by Apoptosis -- I. Introduction -- II. Changes in Light Scatter during Cell Death -- III. Cell Sensitivity to Trypsin and DNase -- IV. Fluorescein Diacetate (FDA) Hydrolysis and PI Exclusion -- V. Rh123 Uptake and PI Exclusion -- VI. PI Exclusion Followed by Counterstaining with Hoechst 33342 -- VII. Hoechst 33342 Active Uptake and PI Exclusion -- VIII. Controlled Extraction of Low MW DNA from Apoptotic Cells -- IX. Sensitivity of DNA in Situ to Denaturation -- X. Detection of DNA Strand Breaks in Apoptotic Cells -- XI. A Selective Procedure for DNA Extraction from Apoptotic Cells Applicable to Gel Electrophoresis and Flow Cytometry -- XII. Comparison of the Methods: Confirmation of the Apoptotic Mode of Cell Death -- References -- Chapter 3. Cell Preparation for the Identification of Leukocytes -- I. Introduction -- II. Antibodies -- III. Titering Antibodies -- IV. Third Color Reagents -- V. Cell Preparation -- VI. Labeling Procedures -- VII. Lysing Erythrocytes -- VIII. Detecting Dead Cells -- IX. Fixation -- X. Solutions and Reagents -- References -- Chapter 4. Multiparameter Analysis of Leukocytes by Flow Cytometry -- I. Introduction -- II. Correlated List-Mode Data -- III. Single-Label Analysis -- IV. Optical Filtration -- V. Compensation -- VI. Two-Color Fluorescence -- VII. Three-Color Immunophenotyping -- VIII. Fixing Cells -- IX. Detecting Dead Cells -- X. Summary -- References. , Chapter 5. Immunophenotyping Using Fixed Cells -- I. Introduction -- II. Application -- III. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Results and Discussion -- References -- Chapter 6. Simultaneous DNA Content and Cell-Surface Immunofluorescence Analysis -- I. Introduction -- II. Materials and Methods -- III. Instruments -- IV. Critical Aspects of the Method -- V. Results -- VI. Applications and Future Directions -- References -- Chapter 7. Flow Cytometry Crossmatching for Solid Organ Transplantation -- I. Introduction -- II. Applications -- Ill. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Standards -- VII. Instruments -- VIII. Results and Discussion -- References -- Chapter 8. Cell Membrane Potential Analysis -- I. Introduction -- II. Materials and Methods -- III. Improving Cytometry of Membrane Potentials -- References -- Chapter 9. Measurement of Intracellular pH -- I. Introduction -- II. Application -- Ill. Materials -- IV. Cell Preparation and Staining: BCECF or SNARF -- V. lnstruments -- VI. Critical Aspects -- VII. Results and Discussion -- References -- Chapter 10. Intracellular Ionized Calcium -- I. Introduction -- II. Flow Cytometric Assay With Indo-1 -- III. Use of Flow Cytometry and Fluo-3 to Measure [Ca2+]i -- IV. Use of Flow Cytometry and Fluo-3/Fura-Red for Ratiometric Analysis of [Ca2+]i -- V. Cell Conjugate Assays Combined with Calcium Analysis and Flow Cytometry -- VI. Pitfalls and Critical Aspects -- VII. Limitations -- VIII. Results -- References -- Chapter 11. Cellular Protein Content Measurements -- I. lntroduction -- II. Applications -- III. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results and Discussion -- References. , Chapter 12. Lysosomal Proton Pump Activity: Supravital Cell Staining with Acridine Orange Differentiates Leukocyte Subpopulations -- I. Introduction -- II. Applications -- III. Materials -- IV. Staining Procedures -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- References -- Chapter 13. Staining of DNA in Live and Fixed Cells -- I. Introduction -- II. Applications with DNA-Specific Fluorochromes -- III. Materials -- IV. Cell Preparation and Fixation -- V. Cell Staining -- VI. Critical Aspects -- VII. Controls and Standards -- VIII. Instruments -- IX. Results and Discussion -- References -- Chapter 14. High-Resolution Analysis of Nuclear DNA Employing the Fluorochrome DAPI -- I. Introduction -- II. Applications -- III. Materials -- IV. Cell Separation and Fixation Method -- V. Staining Procedure -- VI. Instrument Setup -- VII. Standards -- VIII. Results and Discussion -- References -- Chapter 15. 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 -- References -- Chapter 16. DNA Analysis from Paraffin-Embedded Blocks -- I. Introduction -- II. Application -- III. Methods -- IV. Critical Aspects of Technique -- V. Alternative Methods for Sample Preparation -- References -- Chapter 17. Controls, Standards, and Histogram Interpretation in DNA Flow Cytometry -- I. Introduction -- II. Application -- III. Instrument Quality Control and Standardization -- IV. DNA Histogram Interpretation -- V. Methods -- VI. Summary of Criteria for Interpretation of DNA Histograms -- References -- Chapter 18. DNA Content Histogram and Cell-Cycle Analysis -- I. Introduction -- 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 19. Imniunochemical Quantitation of Bromodeoxyuridine: Application to Cell-Cycle Kinetics -- I. Introduction -- II. Applications -- III. Materials -- IV. Procedures -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- IX. Summary -- References -- Chapter 20. Application and Detection of IdUrd and CldUrd as Two Independent Cell-Cycle Markers -- I. Introduction -- II. Application -- III. Materials -- IV. Methods -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Results -- References -- Chapter 21. Cell-Cycle Analysis Using Continuous Bromodeoxyuridine Labeling and Hoechst 33358-Ethidium Bromide Bivariate Flow Cytometry -- I. Introduction -- II. Application -- III. Materials -- IV. Procedures -- V. Controls and Standards -- VI. Instruments -- VII. Results -- References -- Chapter 22. Detection of BrdUrd-Labeled Cells by Differential Fluorescence Analysis of DNA Fluorochromes: Pulse-Chase and Continuous Labeling Methods -- I. Introduction -- 11. Applications -- 111. Materials -- IV. Cell Preparation and Staining Procedure -- V. Critical Aspects -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- References -- Chapter 23. Analysis of Intracellular Proteins -- I. Introduction -- II. Application -- III. Materials -- IV. Staining Procedures -- V. Critical Aspects -- VI. Controls and Standards -- VII. Instruments -- VIII. Multiparameter Analysis of Intracellular Antigens -- References -- Chapter 24. "Washless" Procedures for Nuclear Antigen Detection -- I. Introduction -- II. Applications -- Ill. Materials, Cell Preparation, and Staining. , IV. Critical Aspects of the Procedure: Controls, Standards, and Instruments -- V. Results -- References -- Chapter 25. Light Scatter of Isolated Cell Nuclei as a Parameter Discriminating the Cell-Cycle Subcompartments -- I. Introduction -- II. Application -- III. Materials -- IV. Critical Aspects of the Procedure -- V. Instruments -- Vl. Results -- VII. Appendix -- References -- Chapter 26. Simultaneous Analysis of Cellular RNA and DNA Content -- I. Introduction -- II. Applications -- III. Materials -- IV. Staining Procedures Employing A O -- V. Staining RNA and DNA with PY and Hoechst 33342 -- VI. Critical Aspects of the Procedures -- VII. Controls and Standards -- VIII. Instruments -- IX. Results -- X. Comparison of the Methods -- References -- Chapter 27. Analysis of DNA Content and Cyclin Protein Expression in Studies of DNA Ploidy, Growth Fraction, Lymphocyte Stimulation, and the Cell Cycle -- I. Introduction -- II. Bivariate Analysis of DNA Content versus Cyclin B1 or Cyclin E Expression -- III. Application of Cyclin B1 Analysis in DNA Ploidy and Chromatin Cycle Estimates -- IV. Lymphocyte Stimulation -- V. Tumor Growth Fraction Analysis -- Vl. Mapping the Cell Cycle: Mechanism of Action of Cytostatic Drugs -- VII. Critical Points of the Procedure -- VIII. Conclusions -- References -- Chapter 28. Oxidative Product Formation Analysis by Flow Cytometry -- I. Introduction -- II. Oxidative Burst -- III. Phagocytosis -- IV. Controls and Standards -- References -- Chapter 29. Flow Cytometric Determination of Cysteine and Serine Proteinase Activities in Living Cells with Rhodamine 110 Substrates -- I. Introduction -- II. Materials and Methods -- III. Results -- IV. Discussion -- References -- Chapter 30. Leucine Aminopeptidase Activity by Flow Cytometry -- I. Introduction -- II. Application -- III. Materials -- IV. Methods. , V. Critical Aspects of the Procedure.