Elsevier S & T

Cytometry, Part A

Wilson, Leslie

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Table of contents

Cytometry, Part a, 3rd EditionCover
Cover
Cover
CONTENTSv
CONTENTSv
Contributorsxvii
Contributorsxvii
Preface to the Third Editionxxi
Preface to the Third Editionxxi
Preface to the Second Editionxxv
Preface to the Second Editionxxv
Preface to the First Editionxxix
Preface to the First Editionxxix
Part I: Principles of Cytometry and General Methods1
Part I: Principles of Cytometry and General Methods1
Chapter 1. A Brief History of Flow Cytometry and Sorting3
Chapter 1. A Brief History of Flow Cytometry and Sorting3
I. Introduction3
I. Introduction3
II. Instrumentation4
II. Instrumentation4
III. Applications9
III. Applications9
References13
References13
Chapter 2. Principles of Flow Cytometry: An Overview19
Chapter 2. Principles of Flow Cytometry: An Overview19
I. Introduction19
I. Introduction19
II. The Illumination of a Particle20
II. The Illumination of a Particle20
III. Fluidics: Centering Particles in the Illuminating Beam26
III. Fluidics: Centering Particles in the Illuminating Beam26
IV. Collection of Light Signals from Particles33
IV. Collection of Light Signals from Particles33
V. From Light Signals to a Data File37
V. From Light Signals to a Data File37
VI. From Data to Information41
VI. From Data to Information41
VII. Sorting44
VII. Sorting44
VIII. Conclusions48
VIII. Conclusions48
References48
References48
Chapter 3. Laser Scanning Cytometry51
Chapter 3. Laser Scanning Cytometry51
I. Introduction51
I. Introduction51
II. Background52
II. Background52
III. Description of the Instrument55
III. Description of the Instrument55
IV. The Utility and Operational Characteristics of Some Laser Scanning Cytometry List Mode Features62
IV. The Utility and Operational Characteristics of Some Laser Scanning Cytometry List Mode Features62
V. Utility of Solid Phase Cytometry for Cell Preparation80
V. Utility of Solid Phase Cytometry for Cell Preparation80
VI. Future Directions82
VI. Future Directions82
References84
References84
Chapter 4. Principles of Confocal Microscopy89
Chapter 4. Principles of Confocal Microscopy89
I. Brief History of Microscope Development89
I. Brief History of Microscope Development89
II. Development of Confocal Microscopy90
II. Development of Confocal Microscopy90
III. Image Formation in Confocal Microscopy92
III. Image Formation in Confocal Microscopy92
IV. Useful Fluorescent Probes for Confocal Microscopy96
IV. Useful Fluorescent Probes for Confocal Microscopy96
V. Applications of Confocal Microscopy100
V. Applications of Confocal Microscopy100
VI. Conclusions105
VI. Conclusions105
References105
References105
Chapter 5. Optical Measurements in Cytometry: Light Scattering, Extinction, Absorption, and Fluoresc107
Chapter 5. Optical Measurements in Cytometry: Light Scattering, Extinction, Absorption, and Fluoresc107
I. Introduction108
I. Introduction108
II. Signal Processing Tasks in Flow Cytometry: An Overview108
II. Signal Processing Tasks in Flow Cytometry: An Overview108
III. The Optical Signal: Interaction of Light with Cells110
III. The Optical Signal: Interaction of Light with Cells110
IV. Detection: Converting Optical Signals to Current118
IV. Detection: Converting Optical Signals to Current118
V. Electronics: Converting Current to Voltage123
V. Electronics: Converting Current to Voltage123
VI. Fluorescence Compensation and Logarithmic Amplification124
VI. Fluorescence Compensation and Logarithmic Amplification124
VII. Peak Detection, Integration, and Pulse Width Measurement. Triggering127
VII. Peak Detection, Integration, and Pulse Width Measurement. Triggering127
VIII. Measurement Sensitivity: Changing Concepts and the Bottom Line127
VIII. Measurement Sensitivity: Changing Concepts and the Bottom Line127
References129
References129
Chapter 6. Flow Cytometric Fluorescence Lifetime Measurements131
Chapter 6. Flow Cytometric Fluorescence Lifetime Measurements131
I. Introduction131
I. Introduction131
II. Applications of the Technology134
II. Applications of the Technology134
III. Cell Preparation and Staining137
III. Cell Preparation and Staining137
IV. Fluorescence Lifetime Flow Cytometry Instrumentation138
IV. Fluorescence Lifetime Flow Cytometry Instrumentation138
V. Results141
V. Results141
VI. Critical Aspects of the Technology144
VI. Critical Aspects of the Technology144
VII. Future Directions146
VII. Future Directions146
References147
References147
Chapter 7. Principles of Data Acquisition and Display149
Chapter 7. Principles of Data Acquisition and Display149
I. Introduction149
I. Introduction149
II. Pulse Characterization Using Analog and Hybrid Circuits150
II. Pulse Characterization Using Analog and Hybrid Circuits150
III. Analog Data Display154
III. Analog Data Display154
IV. Analog-to-Digital Conversion155
IV. Analog-to-Digital Conversion155
V. Pulse Characterization by Digital Signal Processing159
V. Pulse Characterization by Digital Signal Processing159
VI. Data Storage and Display with Digital Computers161
VI. Data Storage and Display with Digital Computers161
References167
References167
Chapter 8. Time as a Flow Cytometric Parameter169
Chapter 8. Time as a Flow Cytometric Parameter169
I. Introduction169
I. Introduction169
II. Historical Overview171
II. Historical Overview171
III. Sample Mixing and Delivery171
III. Sample Mixing and Delivery171
IV. Data Analysis175
IV. Data Analysis175
V. Applications179
V. Applications179
VI. Conclusions181
VI. Conclusions181
References181
References181
Chapter 9. Protein Labeling with Fluorescent Probes185
Chapter 9. Protein Labeling with Fluorescent Probes185
I. Introduction185
I. Introduction185
II. Labeling of Proteins with Organic Fluorescent Dyes186
II. Labeling of Proteins with Organic Fluorescent Dyes186
III. Labeling of Proteins with Phycobiliproteins194
III. Labeling of Proteins with Phycobiliproteins194
IV. Conclusion202
IV. Conclusion202
References202
References202
Part II: Cell Preparation205
Part II: Cell Preparation205
Chapter 10. Preparation of Cells from Blood207
Chapter 10. Preparation of Cells from Blood207
I. Introduction207
I. Introduction207
II. Collection, Transport, and Storage of Blood208
II. Collection, Transport, and Storage of Blood208
III. Fixation and Preservation210
III. Fixation and Preservation210
IV. Separation of Erythrocytes from Leukocytes210
IV. Separation of Erythrocytes from Leukocytes210
V. Assessment of Cell Viability211
V. Assessment of Cell Viability211
VI. Staining212
VI. Staining212
VII. Summary212
VII. Summary212
Appendix 1213
Appendix 1213
Appendix 2214
Appendix 2214
Appendix 3214
Appendix 3214
References214
References214
Chapter 11. Cell Preparation for the Identification of Leukocytes217
Chapter 11. Cell Preparation for the Identification of Leukocytes217
I. Introduction218
I. Introduction218
II. Antibodies219
II. Antibodies219
III. Tandem Fluorochromes234
III. Tandem Fluorochromes234
IV. Cell Preparation and Staining Procedures237
IV. Cell Preparation and Staining Procedures237
V. Titering Antibodies244
V. Titering Antibodies244
VI. Solutions and Reagents249
VI. Solutions and Reagents249
References250
References250
Chapter 12. Strategies for Cell Permeabilization and Fixation in Detecting Surface and Intracellular253
Chapter 12. Strategies for Cell Permeabilization and Fixation in Detecting Surface and Intracellular253
I. Introduction253
I. Introduction253
II. Application254
II. Application254
III. Materials and Methods262
III. Materials and Methods262
IV. Concluding Remarks266
IV. Concluding Remarks266
References266
References266
Part III: Standardization, Quality Assurance269
Part III: Standardization, Quality Assurance269
Chapter 13. Stoichiometry of Immunocytochemical Staining Reactions271
Chapter 13. Stoichiometry of Immunocytochemical Staining Reactions271
I. Introduction271
I. Introduction271
II. Structure of Immunoglobulin G273
II. Structure of Immunoglobulin G273
III. Cell Structure274
III. Cell Structure274
IV. Permeabilized Cell Structure274
IV. Permeabilized Cell Structure274
V. Antibody–Antigen Reactions281
V. Antibody–Antigen Reactions281
VI. Multiparametric Analyses292
VI. Multiparametric Analyses292
VII. Summary294
VII. Summary294
References295
References295
Chapter 14. Standardization and Quantitation in Flow Cytometry299
Chapter 14. Standardization and Quantitation in Flow Cytometry299
I. Introduction300
I. Introduction300
II. General Issues300
II. General Issues300
III. Performance Characteristics„Dynamic Range, Linearity, Resolution, and Sensitivity302
III. Performance Characteristics„Dynamic Range, Linearity, Resolution, and Sensitivity302
IV. Standardization and Calibration of Common Cytometry Measurements312
IV. Standardization and Calibration of Common Cytometry Measurements312
V. Examples of Applications Using Calibrated Measurements330
V. Examples of Applications Using Calibrated Measurements330
VI. Issues in Quantitation of Fluorochromes and Other Molecules332
VI. Issues in Quantitation of Fluorochromes and Other Molecules332
References334
References334
Part IV: Cell Proliferation341
Part IV: Cell Proliferation341
Chapter 15. Methods to Identify Mitotic Cells by Flow Cytometry343
Chapter 15. Methods to Identify Mitotic Cells by Flow Cytometry343
I. Introduction343
I. Introduction343
II. Materials345
II. Materials345
III. Cell Preparation and Staining345
III. Cell Preparation and Staining345
IV. Instruments346
IV. Instruments346
V. Critical Aspects of the Procedure346
V. Critical Aspects of the Procedure346
VI. Results and Discussion347
VI. Results and Discussion347
VII. Comparison of Anti-H3-P Monoclonal Antibody with Other Markers of Mitotic Cells350
VII. Comparison of Anti-H3-P Monoclonal Antibody with Other Markers of Mitotic Cells350
References353
References353
Chapter 16. Cell Cycle Kinetics Estimated by Analysis of Bromodeoxyuridine Incorporation355
Chapter 16. Cell Cycle Kinetics Estimated by Analysis of Bromodeoxyuridine Incorporation355
I. Introduction355
I. Introduction355
II. Applications357
II. Applications357
III. Materials358
III. Materials358
IV. Methods360
IV. Methods360
V. Critical Aspects of the Procedure369
V. Critical Aspects of the Procedure369
References372
References372
Chapter 17. Flow Cytometric Analysis of Cell Division History Using Dilution of Carboxyfluorescein D375
Chapter 17. Flow Cytometric Analysis of Cell Division History Using Dilution of Carboxyfluorescein D375
I. Introduction and Background376
I. Introduction and Background376
II. Reagents and Solutions378
II. Reagents and Solutions378
III. Preparation and Labeling of Cells378
III. Preparation and Labeling of Cells378
IV. Gathering of Information Concurrent with Division381
IV. Gathering of Information Concurrent with Division381
V. Analysis of Data386
V. Analysis of Data386
VI. Application of Carboxyfluorescein Diacetate Succinimidyl Ester to in Vitro Culture of Lymphocyt390
VI. Application of Carboxyfluorescein Diacetate Succinimidyl Ester to in Vitro Culture of Lymphocyt390
VII. Monitoring Lymphocyte Responses in Vivo392
VII. Monitoring Lymphocyte Responses in Vivo392
VIII. Antigen Receptor Transgenic Models395
VIII. Antigen Receptor Transgenic Models395
References397
References397
Chapter 18. Antibodies against the Ki-67 Protein: Assessment of the Growth Fraction and Tools for Ce399
Chapter 18. Antibodies against the Ki-67 Protein: Assessment of the Growth Fraction and Tools for Ce399
I. Introduction399
I. Introduction399
II. Application401
II. Application401
III. Materials and Methods402
III. Materials and Methods402
IV. Critical Aspects408
IV. Critical Aspects408
V. Controls and Standards409
V. Controls and Standards409
VI. Examples of Results410
VI. Examples of Results410
References414
References414
Chapter 19. Detection of Proliferating Cell Nuclear Antigen419
Chapter 19. Detection of Proliferating Cell Nuclear Antigen419
I. Introduction419
I. Introduction419
II. Molecular Biology of Proliferating Cell Nuclear Antigen420
II. Molecular Biology of Proliferating Cell Nuclear Antigen420
III. Methods for Immunochemical Detection and Quantification of Proliferating Cell Nuclear Antigen421
III. Methods for Immunochemical Detection and Quantification of Proliferating Cell Nuclear Antigen421
IV. Results of Cytometric Analysis of Proliferating Cell Nuclear Antigen Expression423
IV. Results of Cytometric Analysis of Proliferating Cell Nuclear Antigen Expression423
V. Applications in Toxicology, Pathology, and Oncology426
V. Applications in Toxicology, Pathology, and Oncology426
References428
References428
Chapter 20. Lymphocyte Activation Associated Antigens433
Chapter 20. Lymphocyte Activation Associated Antigens433
I. Introduction433
I. Introduction433
II. Methodological Aspects437
II. Methodological Aspects437
III. To Flow or Not to Flow for Assessing Lymphocyte Activation/ Proliferation? And, If Yes, How Re447
III. To Flow or Not to Flow for Assessing Lymphocyte Activation/ Proliferation? And, If Yes, How Re447
IV. Additional Approaches453
IV. Additional Approaches453
V. Concluding Remarks455
V. Concluding Remarks455
References457
References457
Part V: Cell Death/Apoptosis465
Part V: Cell Death/Apoptosis465
Chapter 21. Analysis of Mitochondria during Cell Death467
Chapter 21. Analysis of Mitochondria during Cell Death467
I. Introduction467
I. Introduction467
II. Scientific Background468
II. Scientific Background468
III. Apoptosis and Mitochondria470
III. Apoptosis and Mitochondria470
IV. Method472
IV. Method472
V. Results473
V. Results473
VI. Pitfalls and Misinterpretation of the Data476
VI. Pitfalls and Misinterpretation of the Data476
VII. Comparison with Other Methods477
VII. Comparison with Other Methods477
VIII. Reviews of the Applications478
VIII. Reviews of the Applications478
IX. Biological and Biomedical Information479
IX. Biological and Biomedical Information479
X. Future Directions480
X. Future Directions480
References481
References481
Chapter 22. Cytometry of Caspases487
Chapter 22. Cytometry of Caspases487
I. Introduction487
I. Introduction487
II. Materials and Methods: Caspase Peptide Inhibitors and Methods to Monitor Responses490
II. Materials and Methods: Caspase Peptide Inhibitors and Methods to Monitor Responses490
III. Results and Discussion497
III. Results and Discussion497
References502
References502
Chapter 23. Analysis of Apoptosis in Plant Cells505
Chapter 23. Analysis of Apoptosis in Plant Cells505
I. Introduction505
I. Introduction505
II. Apoptosis in Plants506
II. Apoptosis in Plants506
III. Problems Associated with Analyzing Plant Cells Using Flow Cytometry509
III. Problems Associated with Analyzing Plant Cells Using Flow Cytometry509
IV. Morphological Changes of Plant Cells512
IV. Morphological Changes of Plant Cells512
V. Physiological Changes during Apoptosis520
V. Physiological Changes during Apoptosis520
VI. Conclusion524
VI. Conclusion524
References524
References524
Chapter 24. Difficulties and Pitfalls in Analysis of Apoptosis527
Chapter 24. Difficulties and Pitfalls in Analysis of Apoptosis527
I. Introduction527
I. Introduction527
II. Apoptotic Index May Not Be Correlated with Incidence of Cell Death529
II. Apoptotic Index May Not Be Correlated with Incidence of Cell Death529
III. Dif.culties in Estimating Frequency of Apoptosis by Analysis of DNA Fragmentation531
III. Dif.culties in Estimating Frequency of Apoptosis by Analysis of DNA Fragmentation531
IV. The Lack of Evidence Is Not Evidence for the Lack of Apoptosis532
IV. The Lack of Evidence Is Not Evidence for the Lack of Apoptosis532
V. Misclassification of Apoptotic Bodies or Nuclear Fragments as Single Apoptotic Cells533
V. Misclassification of Apoptotic Bodies or Nuclear Fragments as Single Apoptotic Cells533
VI. Apoptosis versus Necrosis versus ‘‘Necrotic Stage‘‘ of Apoptosis535
VI. Apoptosis versus Necrosis versus ‘‘Necrotic Stage‘‘ of Apoptosis535
VII. Selective Loss of Apoptotic Cells during Sample Preparation537
VII. Selective Loss of Apoptotic Cells during Sample Preparation537
VIII. Live Cells Engulfing Apoptotic Bodies Masquerade as Apoptotic Cells538
VIII. Live Cells Engulfing Apoptotic Bodies Masquerade as Apoptotic Cells538
IX. The Problems with Commercial Kits and Reagents538
IX. The Problems with Commercial Kits and Reagents538
X. Cell Morphology Is Still the Gold Standard for Identification of Apoptotic Cells539
X. Cell Morphology Is Still the Gold Standard for Identification of Apoptotic Cells539
XI. Laser Scanning Cytometry: Have Your Cake and Eat It Too541
XI. Laser Scanning Cytometry: Have Your Cake and Eat It Too541
References544
References544
Part VI: Cell–Cell, Cell–Environment Interactions547
Part VI: Cell–Cell, Cell–Environment Interactions547
Chapter 25. Analysis of Cell Migration549
Chapter 25. Analysis of Cell Migration549
I. Introduction and Application549
I. Introduction and Application549
II. General Strategies to Measure Cell Migration550
II. General Strategies to Measure Cell Migration550
References558
References558
Chapter 26. Three-Dimensional Extracellular Matrix Substrates for Cell Culture561
Chapter 26. Three-Dimensional Extracellular Matrix Substrates for Cell Culture561
I. Introduction561
I. Introduction561
II. Application563
II. Application563
III. Methods565
III. Methods565
IV. Application of Intestinal Submucosa as a Three-Dimensional Extracellular Matrix Substrate572
IV. Application of Intestinal Submucosa as a Three-Dimensional Extracellular Matrix Substrate572
V. Summary576
V. Summary576
References578
References578
Chapter 27. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix–Cell Intera583
Chapter 27. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix–Cell Intera583
I. Introduction583
I. Introduction583
II. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix–Cell Interactions:584
II. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix–Cell Interactions:584
III. Three-Dimensional Microscopy of Living Systems: Extracellular Matrix and Extracellular Matrix??587
III. Three-Dimensional Microscopy of Living Systems: Extracellular Matrix and Extracellular Matrix??587
IV. Summary593
IV. Summary593
References596
References596
Chapter 28. Cytometric Analysis of Cell Contact and Adhesion599
Chapter 28. Cytometric Analysis of Cell Contact and Adhesion599
I. Introduction and Application599
I. Introduction and Application599
II. General Strategies to Measure Cell–Cell Adhesions600
II. General Strategies to Measure Cell–Cell Adhesions600
III. General Strategies to Measure Cell–Ligand Adhesions606
III. General Strategies to Measure Cell–Ligand Adhesions606
IV. Specificity of Cell Adhesion608
IV. Specificity of Cell Adhesion608
V. Optimization of Experimental Conditions609
V. Optimization of Experimental Conditions609
References611
References611
Chapter 29. Invadopodia: Unique Methods for Measurement of Extracellular Matrix Degradation in Vitro613
Chapter 29. Invadopodia: Unique Methods for Measurement of Extracellular Matrix Degradation in Vitro613
I. Introduction613
I. Introduction613
II. Invadopodia Activity, a Measurement for Localized Membrane Degradation615
II. Invadopodia Activity, a Measurement for Localized Membrane Degradation615
III. Fluorescent Activated Cell Sorting„Phagocytosis, a Measurement for Internalization of Proteol619
III. Fluorescent Activated Cell Sorting„Phagocytosis, a Measurement for Internalization of Proteol619
IV. Protocols623
IV. Protocols623
References626
References626
Index629
Index629
Volumes in Series645
Volumes in Series645

Book details

Vendor Elsevier S & T
SKU 9780125441667
ISBN-13 9780080522524
Author Wilson, Leslie
Edition 3rd
Category Science
Subject Research & Methodology

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Each chapter presents a detailed background of the described method, its theoretical foundations, and its applicability to different biomedical material. Updated chapters describe either the most popular methods or those processes that have evolved the most since the past edition. Additionally, a large portion of the volume is devoted to clinical cytometry. Particular attention is paid to applications of cytometry in oncology, the most rapidly growing area.

Key Features
* Contains 56 extensive chapters authored by world authorities on cytometry
* Covers a wide range of topics, including principles of cytometry and general methods, cell preparation, tandardization and quality assurance, cell proliferation, apoptosis, cell-cell/cell-environmental interactions, cytogenetics and molecular genetics, cell function and differentiation, experimental and clinical oncology, microorganisms, and infectious diseases
* Describes in-depth the essential methods and scientific principles of flow and laser scanning cytometry and illustrates how they can be applied to the fields of biology and medicine
* Complements the first and second editions on flow cytometry in the Methods in Cell Biology> series and includes new sections on technology principles.

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