Flow Cytometry in Hematopathology (eBook)

Contents 6
Preface to the Second Edition 10
Preface to the First Edition 11
Acknowledgments 13
List of Abbreviations 14
List of Case Studies 16
1 Approach to flow cytometry: General considerations 19
1.1 Reasons for the necessity of proper data analysis 20
1.2 General aspects of FCM data analysis and interpretation 22
1.3 Other applications of FCM in hematopathology 25
1.4 Maturation and differentiation of hematopoietic elements: An overview based on the immunologic markers currently in use in the FCM laboratory 27
2 FCM immunophenotyping and DNA analysis: Practical aspects that can affect data analysis and interpretation 31
2.1 Sample selection 31
2.1.1 Liquid specimens 32
2.1.2 Solid tissue specimens 33
2.2 Preparing nucleated cell suspensions 34
2.3 Cell yield and viability 34
2.4 Sample staining 34
2.4.1 Surface antigens 35
2.4.2 Intracellular antigens 35
2.4.3 DNA content 36
2.5 Data acquisition 37
2.5.1 Calibration 37
2.5.2 Color compensation 37
2.5.3 List mode data collection 38
2.5.4 Exclusion of nonviable cells 39
2.6 Antibody panel design 40
2.6.1 Antibody selection 41
2.6.2 Fluorochrome conjugation 44
2.7 Comprehensive antibody panels 49
2.7.1 Disease-oriented antibody panels 49
2.7.2 Antibody panels oriented by specimen type 50
2.8 Tailored panels and add-on testing 52
2.9 FCM immunophenotyping data representation 55
2.9.1 Analysis panels 55
2.9.2 Color display 56
2.10 Approach to DNA data analysis 59
2.10.1 DNA ploidy 60
2.10.2 S-phase 60
3 FCM data analysis on nearly homogeneous samples 67
3.1 FCM parameters 68
3.1.1 Forward scatter 68
3.1.2 Side scatter 71
3.1.3 Fluorescence 72
3.2 Fluorescence dynamic range 89
3.3 Strategy to the visual review of FCM immunophenotyping data 89
3.4 Common SSC/CD45 patterns 91
3.4.1 Assessment of the blast population 91
3.4.2 Immature neoplastic cells with downregulated CD45 99
3.4.3 SSC/CD45 in mature lymphoid disorders 101
3.5 Other dot plot patterns useful in acute leukemia diagnosis 104
3.5.1 Useful antigenic features in AML 104
3.5.2 Precursor B-ALL versus bone marrow B-cell progenitors 116
3.5.3 Useful antigenic features in precursor T-lymphoma/leukemia 117
3.6 Evaluation of mature lymphoid malignancies 124
3.6.1 Assessment of surface light chain expression 124
3.6.2 Assessment of pan B-cell antigens 126
3.6.3 Useful antigenic features in mature B-cell malignancies 130
3.6.4 Identification of abnormal mature T-cells 149
3.6.5 Useful antigenic features in mature T-cell malignancies 168
3.7 Assessing the biological behavior of mature lymphoid neoplasms 184
3.8 Dot plot patterns in histiocytic proliferations and nonhematopoietic malignancies 190
Plates 192
4 FCM data analysis on heterogeneous specimens 209
4.1 Identifying normal FCM samples 209
4.2 Abnormal heterogeneous samples with a detectable immature neoplastic population 254
4.2.1 Blasts of lymphoid lineage 255
4.2.2 Blasts of myeloid lineage 256
4.3 Minimal residual disease 267
4.4 Abnormal heterogeneous samples with detectable mature neoplastic populations 267
4.4.1 Abnormal mature B-cells 272
4.4.2 Abnormal mature T-cells and NK cells 279
4.4.3 Abnormal plasma cells present 289
4.5 Abnormal blood or bone marrow samples with no detectable neoplastic cells 294
4.5.1 Altered cellular composition and abnormal SSC 296
4.5.2 Abnormal antigenic maturation in myeloid or erythroid precursors 307
4.6 Coexisting malignancies 319
5 FCM interpretation and reporting 322
5.1 Immature hematopoietic malignancies 322
5.1.1 ALL/lymphoblastic lymphoma 323
5.1.2 Myeloid malignancies 325
5.1.3 Acute leukemias with a multilineage antigenic pro.le 333
5.2 Mature lymphoid malignancies 333
5.2.1 B-cell LPD/NHL 335
5.2.2 Plasma cell dyscrasias 351
5.2.3 T-cell LPD/NHL 353
5.3 FCM reporting 366
Suggested reading 367
Using the case studies 373
Limited warranty and disclaimer 373
Appendix: Using the case studies CD-ROM 373
Index 374