Darzynkiewicz / Holden / Telford Recent Advances in Cytometry, Part B

Advances in Applications
5. Auflage 2011
ISBN: 978-0-12-385494-0
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, Band Volume 103, 400 Seiten

Reihe: Methods in Cell Biology

Cytometry is one of the most rapidly growing methodologies available for basic cell and molecular biology, cytogenetics, immunology, oncology, environmental sciences and also various fields of clinical medicine. This new edition, split into 2 Parts, is an almost completely new book, with nearly all of the chapters devoted to new topics. Like the previous volumes on cytometry published as part of the Methods in Cell Biology series, it provides a comprehensive description of particular cytometric methods and reviews their applications. Chapters present the theoretical foundations of the described methods, their applicability in experimental laboratory and clinical settings, and describes common traps and pitfalls such as problems with data interpretation, comparison with alternative assays, and choosing the optimal assay. - Comprehensive presentation of cytometric methods covering theoretical applications, applicability, potential pitfalls, and comparisions to alternative assays - Discusses many new assays developed since the previous edition - Presents recent developments in cytometric intrumentation/technology

Autoren/Hrsg.

Darzynkiewicz, Zbigniew

Holden, Elena

Telford, William

Wlodkowic, Donald

Inhaltsverzeichnis

1;Cover;1
2;Methods in Cell Biology Volume 103;2
3;Copyright;5
4;Contents;6
5;Contributors;12
6;Preface To Fifth Edition;16
7;Section - 1 - New applications in cell biology;20
7.1;Chapter - 1 - Recent Advances in Cytometry Applications: Preclinical, Clinical, and•Cell•Biology;22
7.1.1;Abstract;22
7.1.2;Preclinical and Clinical Applications;23
7.1.3;Cell Biology and Cell Transplantation Therapy;30
7.1.4;Acknowledgment;33
7.1.5;References;33
7.2;Chapter - 2 - Detection of Hematopoietic Stem Cells by Flow Cytometry;40
7.2.1;Abstract;40
7.2.2;Introduction;41
7.2.3;The side population is highly enriched in hematopoietic stem cells;43
7.2.4;Surface markers for LT-HSC purification;44
7.2.5;References;47
7.3;Chapter - 3 - Identification of Very Small Embryonic/Epiblast-Like Stem Cells (VSELs) Circulating in Peripheral Blood During Organ/Tissue Injuries;50
7.3.1;Abstract;51
7.3.2;Introduction;51
7.3.3;Background;54
7.3.3.1;Very Small Embryonic-Like Stem Cells (VSELs);54
7.3.3.1.1;Developmental Origin of VSELs;55
7.3.3.1.2;VSELs and Their Unique Molecular Characteristics;56
7.3.3.1.3;Epigenetic Changes of Imprinted Genes that Regulate VSELs Pluripotency;58
7.3.3.2;Mesenchymal Stem Cells (MSCs);59
7.3.3.3;Endothelial Progenitor Cells (EPCs);59
7.3.4;Materials;59
7.3.4.1;Preparation of Peripheral Blood (PB) for Analysis;60
7.3.4.2;Staining of Total PB-Derived Nucleated Cells (TNCs) for Analysis;60
7.3.5;Methods;62
7.3.5.1;Isolation of Total PB-Derived Nucleated Cells (TNCs) by Lysing Red Blood Cells (RBCs);62
7.3.5.2;Staining of PB-Derived TNCs for Flow Cytometric Analysis;62
7.3.5.3;Setting Up Instrument for FACS Analysis;63
7.3.5.4;Identification of VSELs in Human PB and UCB;63
7.3.5.5;Calculation of Absolute Numbers of Target Cells in 1µL of PB63
7.3.5.6;Sorting of Cells;63
7.3.6;Results;64
7.3.7;Critical Aspects of the Methodology;66
7.3.8;Applications;68
7.3.9;Future Directions;69
7.3.10;Acknowledgments;69
7.3.11;References;70
7.4;Chapter - 4 - Apoptosis and Beyond: Cytometry in Studies of Programmed Cell Death;74
7.4.1;Abstract;75
7.4.2;Introduction;75
7.4.3;The Biology of Apoptosis;76
7.4.4;Cytometry in Cell Necrobiology;79
7.4.5;Cytometric Methods to Detect Apoptosis;80
7.4.5.1;Light Scattering Changes in Apoptotic Cells;80
7.4.5.2;Dissipation of Mitochondrial Transmembrane Potential (?&psim)81
7.4.5.3;Activation of Caspases;86
7.4.5.3.1;Fluorochrome-Labeled Inhibitors of Caspases (FLICA);86
7.4.5.3.2;Detection of PARP Cleavage;90
7.4.5.4;Changes in the Plasma Membrane During Apoptosis;91
7.4.5.4.1;Externalization of Phosphatidylserine;91
7.4.5.4.2;Changes in Plasma Membrane Permeability;93
7.4.5.5;Nuclear Hallmarks of Apoptosis;94
7.4.5.5.1;Assessment of Fractional DNA Content (Sub-G1 Fraction);95
7.4.5.5.2;Assessment of DNA Strand Breaks (TUNEL Assay);97
7.4.5.6;SYTO-Based Detection of Apoptosis;98
7.4.6;Time-Window in Measuring Incidence of Apoptosis;100
7.4.7;Multiparameter Detection of Apoptosis: Choosing the Right Method;101
7.4.8;Beyond Apoptosis – Analysis of Alternative Cell Death Modes103
7.4.8.1;Autophagy;103
7.4.8.2;Necrosis;105
7.4.8.3;Cell Senescence;105
7.4.9;Future Outlook;107
7.4.10;Acknowledgements;109
7.4.11;References;109
7.5;Chapter - 5 - Assessment of Oxidative Stress-Induced DNA Damage by Immunoflourescent Analysis of 8-OxodG;118
7.5.1;Abstract;118
7.5.2;Redox Regulation of Cell Fate Signaling;119
7.5.3;8-OxodG as a Marker of Oxidative DNA Damage;120
7.5.3.1;8-OxodG is a Highly Mutagenic DNA Lesion;121
7.5.3.2;8-OxodG as a Biomarker of Cellular Oxidative Damage and its Clinical Relevance;121
7.5.4;Detection of Oxidative DNA Damage Involving 8-OxodG;123
7.5.4.1;Chromatography-Based Direct Assessment of 8-OxodG;124
7.5.4.2;The Indirect Assessment of OxodG Modification of DNA;125
7.5.4.2.1;Indirect Assessment of Oxidative DNA Damage by Immunofluorescence;125
7.5.4.2.1.1;Materials;126
7.5.4.2.1.2;Methodology;126
7.5.4.2.1.3;Results;127
7.5.4.2.1.4;Shortcomings of the Immunofluorescence Assay;128
7.5.5;Concluding Remarks;128
7.5.6;References;128
7.6;Chapter - 6 - Analysis of Individual Molecular Events of•DNA Damage Response by Flow- and•Image-Assisted Cytometry;134
7.6.1;Abstract;135
7.6.2;Introduction;135
7.6.3;Events of the DDR;136
7.6.3.1;Chromatin Decondensation (Relaxation);136
7.6.3.2;Activation of Phosphatidyl Inositol 3'-Kinase-Related Kinases (PIKKs)138
7.6.3.3;Activation of Checkpoint Kinases;139
7.6.3.4;Histone H2AX Phosphorylation;141
7.6.4;Detection of DDR Events by Cytometry;141
7.6.4.1;Chromatin Relaxation (Decondensation);141
7.6.4.2;Recruitment of Mre11;143
7.6.4.3;Immunocytochemical Detection of DDR-Associated ATM, DNA-PKcs and Chk2 Activation, Phosphorylation of p53 and Histone H2AX;144
7.6.5;Application of Cytometry to Detect DDR Induced by.Different Genotoxic Agents;145
7.6.5.1;Assessment of DDR Induced by DNA Topoisomerase Inhibitors;145
7.6.5.2;Induction of DDR by Ionizing Radiation and UV Light;148
7.6.5.3;Induction of DDR by Cigarette Smoke (CS) and Other Environmental Mutagens;150
7.6.5.4;Oxidative DNA Damage;152
7.6.6;Interpretation of Cytometric Data: Role of Image-Assisted Cytometry;154
7.6.7;Role of Microfluidic Lab-on-a-Chip Platforms for DDR Analysis;156
7.6.8;References;158
7.7;Chapter - 7 - Fluorescence-Based Detection and Quantification of Features of Cellular Senescence;168
7.7.1;Abstract;169
7.7.2;Introduction;169
7.7.3;Features Associated with Loss of Reproductive Cell Capability;172
7.7.3.1;Shortened Telomeres in Replicative Senescence;173
7.7.3.2;Telomere Dysfunction-Induced Foci (TIF) in Replicative Senescence;175
7.7.3.3;Loss of the Doubling Capacity of Cells;176
7.7.4;Cellular Hypertrophy;176
7.7.5;Changes Associated with Lysosomes;178
7.7.5.1;Lipofuscin Accumulation;178
7.7.5.1.1;Lipofuscin Autofluorescence in Fluorescence Microscopy;179
7.7.5.1.2;Flow Cytometric Analysis of Cellular Lipofuscin Levels;180
7.7.5.2;Increased Lysosome Content;180
7.7.5.3;Senescence-Associated ß-Galactosidase (SA &beta-Gal) Activity181
7.7.5.3.1;In situ Determination of SA ß-Gal Activity182
7.7.5.3.2;Flow Cytometry for SA ß-Gal-Positive Cells and Quantification of Activity182
7.7.6;Changes Associated with Mitochondria;183
7.7.6.1;Increased Mitochondrial Mass;183
7.7.6.1.1;Quantification of Changes in Mitochondria Mass;184
7.7.6.2;Altered Structural Dynamics of Mitochondria;186
7.7.6.2.1;Visualization of Altered Mitochondria Structure;187
7.7.6.3;Decreased Membrane Potential;187
7.7.6.3.1;Quantitative Comparison of MMP;188
7.7.6.4;Decreased Autophagy;190
7.7.6.4.1;Determination of Autophagy Activity;190
7.7.7;Changes in the Level of Reactive Oxygen Species (ROS);192
7.7.7.1;Changes in Level of Superoxide;193
7.7.7.2;Changes in Level of Hydroxyl Radicals;194
7.7.8;Changes Associated with Nucleus and Chromosomes;195
7.7.9;Use of Flow Cytometry for Analysis of Cellular Senescence;197
7.7.10;Conclusion;197
7.7.11;Acknowledgments;198
7.7.12;References;198
7.8;Chapter - 8 - Measurement of Telomere Length Using•PNA Probe by Cytometry;208
7.8.1;Abstract;208
7.8.2;Introduction to Telomeres;209
7.8.3;Technical Background;210
7.8.4;Methods;212
7.8.4.1;Our Protocol Versus the Original Flow-FISH Protocol;212
7.8.4.1.1;Our Protocol;212
7.8.4.1.2;Original Flow-FISH Protocol;213
7.8.5;Results;214
7.8.6;Critical Aspects of the Flow-FISH Methodology;217
7.8.7;Conclusion and Perspectives;219
7.8.8;References;220
8;Section - 2 - Pre-clinical and clinical applications;222
8.1;Chapter - 9 - Cytometry of Intracellular Signaling: From Laboratory Bench to Clinical Application;224
8.1.1;Abstract;224
8.1.2;Introduction;225
8.1.2.1;Activation States Are Transient;226
8.1.2.2;Most Signaling Pathways Exist in Their Ground State;226
8.1.2.3;Constitutive Activation of Signaling Pathways Can Occur in Leukemia;226
8.1.3;Rationale;228
8.1.4;Methods;231
8.1.4.1;Protocol for Whole Blood Fixation and Red Cell Lysis;231
8.1.4.1.1;Samples;231
8.1.4.1.2;Sample Suitability;231
8.1.4.1.3;Addition of Pathway Activators or Inhibitors;231
8.1.4.1.4;Whole Blood Fixation;232
8.1.4.1.5;Red Cell Lysis;232
8.1.4.1.6;Washing and Alcohol Treatment;232
8.1.4.1.7;Antibody Staining;232
8.1.4.2;Activators and Inhibitors of Signaling Pathways;233
8.1.5;Pathway Activators;233
8.1.6;Pathway Inhibitors;234
8.1.6.1;Freezing Samples;236
8.1.7;Materials;236
8.1.7.1;Fixation and Red Cell Lysis;236
8.1.7.2;Pathway Activators;236
8.1.7.3;Pathway Inhibitors;237
8.1.7.4;Antibodies;237
8.1.8;Discussion;237
8.1.9;Summary;238
8.1.10;References;238
8.2;Chapter - 10 - Immunophenotypic Pattern of Myeloid Populations by Flow Cytometry Analysis;240
8.2.1;Abstract;240
8.2.2;Introduction;241
8.2.3;Granulocytes;241
8.2.4;Basophils;244
8.2.5;Eosinophils;244
8.2.6;Monocytes;245
8.2.7;Erythroid Precursors;246
8.2.8;Megakaryocytes;247
8.2.9;Acute Myeloid Leukemia (AML) with Minimal Differentiation and AML without Maturation;247
8.2.10;AML with Maturation;247
8.2.11;AML with t(8;21);2498.2.12;Acute Promyelocytic Leukemia (APL)249
8.2.13;Acute Myelomonocytic Leukemia (AML-M4);258
8.2.14;Acute Monoblastic Leukemia;259
8.2.15;Acute Erythroid Leukemia (FAB: AML-M6);266
8.2.16;Acute Megakaryoblastic Leukemia (FAB: AML-M7);267
8.2.17;Blastic Plasmacytoid Dendritic Cell Neoplasm;269
8.2.18;Chronic Myeloid Leukemia (CML, BCR–ABL+)270
8.2.19;Granulocytes/Maturing Myeloid Precursors with Dyspoietic Features Associated with Myelodysplastic Syndrome (MDS);270
8.2.20;Chronic Myelomonocytic Leukemia (CMML);281
8.2.21;References;282
8.3;Chapter - 11 - Flow Cytometry-Based Pharmacodynamic Monitoring After Organ Transplantation;286
8.3.1;Abstract;286
8.3.2;Introduction;287
8.3.3;T Cells;288
8.3.4;Regulatory T Cells;292
8.3.5;Dendritic Cells;294
8.3.6;Conclusions;298
8.3.7;References;300
8.4;Chapter - 12 - Clinical Cytometry and Progress in HLA Antibody Detection;304
8.4.1;Abstract;304
8.4.2;Introduction;305
8.4.3;Cell-Based Assays;306
8.4.3.1;Lymphocytotoxicity;306
8.4.3.2;Modified Lymphocytotoxicity Assays;308
8.4.3.3;Flow Cytometry;310
8.4.4;Antigen-Based Assays;311
8.4.4.1;Solid-Phase Immunobinding Assays;311
8.4.4.2;Multiplex Platform for Microparticle Analysis;317
8.4.4.3;Recombinant HLA Antigens;320
8.4.4.4;Determining Antibody Specificity;321
8.4.5;Current Frontiers in Transplant-Related Antibody Testing;323
8.4.5.1;Extended HLA Loci;323
8.4.5.2;Conundrums in HLA Antibody Assignment;323
8.4.5.3;Functional Assessments of HLA Alloantibody by Flow Cytometry;324
8.4.6;Summary;324
8.4.7;References;325
8.5;Chapter - 13 - Clinical Utility of Flow Cytometry in•the•Study of Erythropoiesis and Nonclonal Red Cell Disorders;330
8.5.1;Abstract;330
8.5.2;Keywords;331
8.5.3;Introduction;331
8.5.4;Normal Erythroid Development;331
8.5.4.1;Flow Cytometry for Reticulocyte Enumeration;334
8.5.4.2;Flow Cytometry for the Detection of Fetal–Maternal Hemorrhage337
8.5.4.3;Flow Cytometry for Quantitation of HbF in Sickle Cell Disease;341
8.5.4.4;Flow Cytometry in the Evaluation of Hereditary Spherocytosis;342
8.5.4.5;Measurement of Red Cell Survival and Red Cell Volume;344
8.5.4.6;Cell Cycle Studies;344
8.5.4.7;RBC Surface Abnormalities;345
8.5.5;References;345
8.6;Chapter - 14 - Immunophenotypic Characterization of Bone Marrow Mast Cells in Mastocytosis and•Other Mast Cell Disorders;352
8.6.1;Abstract;353
8.6.2;Introduction;353
8.6.3;Background;356
8.6.4;Methods;358
8.6.4.1;Collection of BM Samples;358
8.6.4.2;Staining of BM Samples;358
8.6.4.3;Panel of Monoclonal Antibodies;359
8.6.4.4;Data Acquisition and Analysis;359
8.6.4.5;Critical Parameters and Troubleshooting;360
8.6.5;Results;362
8.6.5.1;Identification, Enumeration, and Characterization of BM MC by Flow Cytometry;362
8.6.5.2;Immunophenotypic Features of Normal BM MC;362
8.6.5.2.1;BM MC-Committed Precursors and Normal MC Maturation Patterns;363
8.6.5.2.2;Immunophenotype of Mature Resting Normal and Reactive BM MC;363
8.6.5.2.3;Activated MC Immunophenotype;366
8.6.5.3;Immunophenotypic Characteristics of BM MC in Systemic Mastocytosis;367
8.6.5.3.1;Good-Prognosis SM: Indolent Systemic Mastocytosis (ISM) with or without Skin Lesions;368
8.6.5.3.2;Well-Differentiated Systemic Mastocytosis;368
8.6.5.3.3;Poor-Prognosis Systemic Mastocytosis: Aggressive Systemic Mastocytosis and Mast Cell Leukemia;370
8.6.6;Immunophenotypic Analysis of MC from Biological Specimens Other than BM;370
8.6.6.1;Identification of Circulating MC in Peripheral Blood;370
8.6.6.2;Identification of MC in Organic Fluids or Tissues;371
8.6.7;Medical Indications;371
8.6.8;Summary;372
8.6.9;Acknowledgments;372
8.6.10;References;373
9;Index;380
10;Volumes In Series;390
11;Color Plate;400