Elsevier S & T

Cytometry: New Developments: New Developments

Darzynkiewicz, Zbigniew

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Table of contents

Cover
Contentsv
Contributorsxvii
Prefacexxiii
Preface to the Third Editionxxvii
Preface to the Second Editionxxxi
Preface to the First Editionxxxv
Part I: Instrumentation/Fluorochromes1
Chapter 1. Optimization of Emission Optics for Multicolor Flow Cytometry3
I. Introduction3
II. Fluorochromes4
III. Hardware Components Affecting Light Collection8
IV. Maximizing Signal Intensities11
V. Maximizing Signal Collection Quality11
VI. Emission Collection Optimization14
VII. Testing Emission Optics16
References21
Chapter 2. Two-Photon Tissue Cytometry23
I. Introduction23
II. Technical Development of Two-Photon Tissue Cytometry24
III. Experimental Demonstration of Two-Photon Tissue Cytometry32
IV. Future Outlook37
V. Conclusion37
References38
Chapter 3. Confocal Microscopy: Quantitative Analytical Capabilities41
I. Introduction42
II. Cartesian Coordinates, Distance, Size, Shape, and Volume in Confocal Microscopy43
III. Measurements of Fluorescence Intensity in Confocal Microscopy53
IV. Time Resolution in Confocal Microscopy57
V. Confocal Techniques Based on Measurements of Fluorescence Intensity vs Time57
VI. Spectrally Resolved Confocal Microscopy64
VII. Conclusion69
References70
Chapter 4. Surface-Plasmon–Coupled Emission. New Technology for Studying Molecular Processes73
I. Introduction73
II. Overview of Surface Plasmon Resonance75
III. Surface-Plasmon–Coupled Emission80
IV. Experimental Studies of Surface-Plasmon–Coupled Emission83
V. Practical Applications90
VI. Conclusion102
References102
Chapter 5. Cytometry of Fluorescence Resonance Energy Transfer105
I. Introduction105
II. Theory of FRET107
III. Measuring FRET109
IV. Applications132
V. Perspectives142
VI. Conclusions145
References146
Chapter 6. The Rainbow of Fluorescent Proteins153
I. The Fluorescent Proteins153
II. Flow Analysis Using Fluorescent Proteins159
III. Flow Sorting Using Fluorescent Proteins163
IV. Conclusion164
References165
Chapter 7. Labeling Cellular Targets with Semiconductor Quantum Dot Conjugates171
I. Introduction171
II. Selection of QDs and Their Conjugates172
III. Labeling of Fixed Cells for Fluorescence Microscopy173
IV. Measurements on Living Cells Using QD Conjugates179
V. Conclusion181
References182
Chapter 8. Next-Generation Laser Scanning Cytometry185
I. Introduction186
II. LSC-Hybrid Technology186
III. Description of the LSC Platform188
IV. Selected Applications203
V. Utility of Solid-Phase Imaging Cytometry in Life Science Research and Drug Discovery217
References217
Part II: General Techniques219
Chapter 9. Biohazard Sorting221
I. Introduction221
II. Critical Aspects of the Procedure222
III. Applications and Future Directions238
References239
Chapter 10. Guidelines for the Presentation of Flow Cytometric Data241
I. Introduction241
II. General Principles of Graphical Presentation243
III. Statistics251
IV. Subset Analysis253
V. Conclusion256
References256
Chapter 11. Mechanism of Antitumor Drug Action Assessed by Cytometry257
I. Introduction258
II. Choice of a Model System258
III. Drug Dose and Length of Exposure263
IV. Single-Parameter DNA Histogram Analysis265
V. Multiparameter Approaches274
VI. Data Presentation298
VII. Summary and Future Directions299
References301
Chapter 12. Cytometric Methods to Detect Apoptosis307
I. Introduction308
II. Physical and Molecular Features of Cells Dying by Apoptosis or Necrosis310
III. Different Time-Windows for Detection of Apoptosis: Possible Source of Error in Measurement of I316
IV. Light-Scattering Properties of Cells Dying by Apoptosis317
V. Mitochondrial Transmembrane Potential318
VI. Activation of Caspases321
VII. Annexin V Binding325
VIII. DNA Fragmentation326
IX. Susceptibility of DNA to Denaturation329
X. Activation of ‘‘Tissue’’ Transglutaminase331
XI. Measuring Incidence of Apoptosis: Which Method to Choose?332
References335
Chapter 13. Real-Time Analysis of Apoptosis In Vivo343
I. Introduction343
II. In Vivo Membrane Markers344
III. A New Generation of Caspase Markers348
IV. Conclusion351
References351
Chapter 14. Detection of DNA Damage in Individual Cells by Analysis of Histone H2AX Phosphorylation355
I. Introduction355
II. Considerations in the Use of yH2AX as a Measure of Double-Stranded Breaks356
III. Methods of Analysis359
IV. Typical Results363
V. Possible Applications370
VI. Conclusion371
References371
Chapter 15. Cytometry of Freshwater Phytoplankton375
I. Introduction376
II. Background378
III. Methods387
IV. Results393
V. Limitations and Applications399
Future Directions403
References403
Chapter 16. Multiplexed Microsphere Assays for Protein and DNA Binding Reactions409
I. Introduction410
II. Technology and Instrumentation411
III. Methods415
IV. Results418
V. Software420
VI. Critical Aspects of the Methodology421
VII. Comparison with Other Methods424
VIII. Future Directions425
References426
Part III: Immunology/T-Cell Responses431
Chapter 17. Flow Cytometry Applications of MHC Tetramers433
I. Introduction433
II. Materials and Methods436
III. Analyzing Tetramer Data444
IV. Combining Tetramer Staining with Functional Assays446
V. Conclusion447
References448
Chapter 18. Use of Peptides and Peptide Libraries as T-Cell Stimulants in Flow Cytometric Studies453
I. Introduction454
II. Background455
III. Results/Discussion of the Literature459
IV. Technical Considerations464
V. Specific Applications467
VI. Potential Clinical Use471
VII. Protocols473
References475
Chapter 19. Flow Cytometric Analysis of Human Antigen-Specific T-Cell Proliferation481
I. Introduction and Background482
II. Methods483
III. Results484
IV. Critical Aspects of the Methodology487
V. Pitfalls and Misinterpretation of the Data490
VI. Future Directions491
References492
Chapter 20. Detection of T-Cell Degranulation: CD107a and b497
I. Introduction: Methods to Assess CD8+ T-Cell Function497
II. Materials and Methods500
III. Results504
IV. Critical Aspects505
V. Applications508
References510
Chapter 21. T-Cell Responses to Cancer513
I. Introduction514
II. Methods to Identify and Enumerate TAA-Specific T Cells516
III. Important Considerations520
IV. Methods to Further Analyze In Vivo Biology of Antitumor T Cells522
V. Novel Issues and Methods524
VI. Conclusion527
References528
Part IV: Multi-Color Immunophenotyping533
Chapter 22. Multicolor Flow Cytometric Analysis in SIV-Infected Rhesus Macaque535
I. Introduction536
II. Methods537
III. Flow Cytometry Instrumentation and Software538
IV. Instrument Setup539
V. Critical Aspects of the Technology542
VI. Application of 10-Color Analysis to SIV Infection548
VII. Conclusion554
References555
Chapter 23. Multicolor Immunophenotyping: Human Immune System Hematopoiesis559
I. Introduction559
II. Methodology560
III. Normal Immunophenotypic Patterns of Maturation561
IV. Abnormal Immunophenotypic Patterns of Maturation573
References575
Chapter 24. Multicolor Immunophenotyping: Human Mature Immune System577
I. Introduction577
II. Choice of Markers578
III. Staining Panels: Matching Fluorochrome with Cell Marker586
IV. Conclusion592
References593
Chapter 25. Differential Diagnosis of T-Cell Lymphoproliferative Disorders by Flow Cytometry Multico595
I. Introduction596
II. Materials596
III. Methods597
IV. Identification of Abnormal T-Cell Population by Flow Cytometry598
V. Precursor T Lymphoblastic Lymphoma/Leukemia601
VI. Peripheral (Mature/Post-thymic) Lymphoma Versus Precursor T-Lymphoblastic Lymphoma/Leukemia601
VII. Thymocytes from Thymic Hyperplasia/Thymoma Versus Precursor T-Lymphoblastic Lymphoma/Leukemia604
VIII. Mature T-Cell Lymphoproliferative Disorders604
IX. CD117 Expression in T-Cell Lymphoproliferations616
X. Blastic NK Cell Lymphoma (DC-2 Acute Leukemia)618
XI. Conclusion618
References619
Chapter 26. Isolation and Immunophenotyping of Human and Rhesus Macaque Dendritic Cells623
I. Introduction623
II. In Vitro Differentiation of Monocyte-Derived Dendritic Cells625
III. Detection and Sorting Procedures of Subsets of Dendritic Cells625
IV. Culture and Activation of Dendritic Cells635
References637
Chapter 27. B-Cell Immunophenotyping643
I. Introduction643
II. Immunophenotyping of B-Cell Developmental Stages in the Bone Marrow644
III. Peripheral B-Cell Populations649
IV. Antigen-Induced B-Cell Subsets655
V. Antigen-Specific B Cells658
References660
Part V: Other Topics in Immunology663
Chapter 28. Flow Cytometry Immunophenotypic Characteristics of Monocytic Population in Acute Monocyt665
I. Introduction666
II. Materials666
III. Methods667
IV. Acute Monocytic Leukemia668
V. Chronic Myelomonocytic Leukemia668
VI. Acute Myelomonocytic Leukemia668
VII. Differential Diagnosis670
VIII. Conclusion676
References676
Chapter 29. Phagocyte Function679
I. Introduction680
II. Background690
III. Methods694
IV. Results697
V. Pitfalls and Misinterpretation of the Data698
VI. Comparison with Other Methods700
VII. Applications and Biomedical Information701
VIII. Future Directions703
References703
Chapter 30. Neutralizing Antibody Quantification by Flow Cytometry709
I. Introduction709
II. Background710
III. Results712
IV. Discussion and Key Points714
References715
Part VI: Cytogenetics717
Chapter 31. Telomere Length Measurements Using Fluorescence In Situ Hybridization and Flow Cytometry719
I. Introduction720
II. Background725
III. Methods727
IV. Results739
V. Critical Aspects of Methodology742
VI. Pitfalls and Misinterpretation of Data744
VII. Comparison with Other Methods745
VIII. Applications746
IX. Future Directions748
References749
Chapter 32. Detecting Copy Number Changes in Genomic DNA: MAPH and MLPA751
I. Introduction751
II. Reactions752
III. Analysis756
IV. Applications762
V. Future Possibilities764
References766
Chapter 33. Genomic Array Technology769
I. Introduction769
II. Construction of Genomic Clone Arrays Using DOP-PCR Amplification770
III. Validation of Array Performance and Data Analysis771
IV. Array CGH776
V. Array CGH for Cytogenetic Analyses778
VI. Array Painting779
VII. Application of Array CGH and Array Painting for Complete Cytogenetic Analyses780
VIII. ChIP on Genomic Clone Arrays782
IX. Conclusion782
References784
Chapter 34. Prospects for In Situ Analyses of Individual and Complexes of DNA, RNA, and Protein Mole787
I. Need for Single-Molecule In Situ Analyses787
II. Problem of Specificity and Sensitivity788
III. Single-or Dual-Recognition Assays789
IV. Some Possible Assay Formats790
V. Specific Detection of Macromolecules followed by Selective In Situ Amplification794
VI. Future Directions795
References795
Chapter 35. The Use of Subtelomeric Probes to Study Mental Retardation799
I. Introduction800
II. Background802
III. Methods806
IV. Results815
V. Critical Aspects of the Methodologies815
VI. Limitations, Pitfalls, and Misinterpretation of Data820
VII. Comparison with Other Methods824
VIII. Applications825
IX. Future Directions826
References828
Index833
Volumes in Series869

Book details

Vendor Elsevier S & T
SKU 9780125641708
ISBN-13 9780080496603
Author Darzynkiewicz, Zbigniew
Edition 4th
Category Science
Subject Cell Biology

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The chapters in CYTOMETRY MCB volumes, including this 4th Edition, provide comprehensive description of particular cytometric methods and review their applications. Some chapters also describe new instrumentation and provide fundamental information on use of new fluorescent probes and on data analysis. Although the term "edition" suggests the update of earlier volumes, in fact, nearly all chapters of the 4th Edition are devoted to new topics. The authors were invited to present not only technical protocols, such as available in other methodology books that specialize in the protocol format, but also to discuss the aspects of the methodology that generally are not included in the protocols. Many chapters, thus, present the theoretical foundations of the described methods, their applicability in experimental laboratory and clinical setting, common traps and pitfalls, problems with data interpretation, comparison with alternative assays, choice of the optimal assay, etc. Some chapters review applications of cytometry and complementary methodologies to particular biological problems or clinical tasks.

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

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