Flow Cytometry in Hematopathology : A Visual Approach to Data Analysis and Interpretation

Preface ix Acknowledgments xi List of Abbreviations xiii Approach to flow cytometry: General considerations Reasons for the necessity of proper data analysis 1(3) The pitfalls of the FCM data format of ``percent positive'' per antibody tested 2(2) General aspects of FCM data analysis and interpretation 4(3) Other applications of FCM in hematopathology 7(1) Overview of lineage-associated markers 7(2) FCM immunophenotyping and DNA analysis: Practical aspects that can affect data analysis and interpretation Sample selection 9(2) Liquid specimens 9(1) Solid tissue specimens 10(1) Preparing nucleated cell suspensions 11(1) Cell yield and viability 12(1) Sample staining 12(2) Surface antigens 12(1) Intracellular antigens 13(1) DNA content 13(1) Data acquisition 14(2) Calibration 14(1) Color compensation 15(1) List mode data collection 15(1) Exclusion of nonviable cells 16(1) Antibody panel design 16(6) Antibody selection 18(1) Anti-light chain antibodies 18(1) Fluorochrome conjugation 19(3) Comprehensive antibody panels 22(3) Disease-oriented antibody panels 22(1) Antibody panels oriented by specimen type 23(2) Tailored panels and add-on testing 25(2) Minimal residual disease 27(1) FCM immunophenotyping data representation 27(5) Analysis panels 28(1) Color display 28(4) Approach to DNA data analysis 32(6) DNA ploidy 32(1) S-phase 33(5) FCM data analysis on nearly homogeneous samples FCM parameters 38(16) Forward scatter 38(4) Side scatter 42(2) Fluorescence 44(3) Heterogeneous fluorescence intensity (bimodal, variable) 47(7) Fluorescence dynamic range 54(1) Strategy to the visual review of FCM immunophenotyping data 54(2) Common SSC/CD45 patterns 56(9) Assessment of the blast population 56(7) Immature neoplastic cells with downregulated CD45 63(1) SSC/CD45 in mature lymphoid disorders 64(1) Other dot plot patterns useful in acute leukemia diagnosis 65(12) Useful antigenic features in AML 66(4) Myeloid phenotypic abnormalities and MRD detection 70(4) Precursor B-ALL vs bone marrow precursor B-cells (hematogones) 74(3) Evaluation of mature lymphoid malignancies 77(32) Assessment of surface light chain expression 77(2) Assessment of pan B-cell antigens 79(4) Useful antigenic features in mature B-cell malignancies 83(1) CD10 expression: Follicular center cell lymphomas 83(3) Pattern of CD20 and CD11c coexpression 86(9) CD5 expression 95(1) Aberrant B-cell profile 96(1) Identification of abnormal T-cells 96(5) Useful antigenic features in mature T-cell malignancies 101(8) Dot plot patterns in histiocytic proliferations and nonhematopoietic malignancies 109(6) FCM data analysis on heterogeneous specimens Identifying normal FCM samples 115(24) Benign/reactive solid lymphoid tissue (e.g., lymph nodes, tonsils) 115(1) Pattern of CD10/CD20 coexpression. Distinction between FRFH and FCC lymphoma 116(7) Normal peripheral blood and normal bone marrow 123(2) Blast region 125(4) Bone marrow B-cell precursors (hematogones) 129(2) Lymphocytes 131(3) Monocytes 134(1) Plasma cells 134(2) Erythroid precursors 136(2) Maturing myeloid cells 138(1) Abnormal samples with a detectable immature neoplastic population 139(13) Blasts of lymphoid lineage 140(1) Blasts of myeloid lineage 141(2) AML 143(3) High-grade MDS and MPD with increased blasts 146(6) Minimal residual disease 152(2) Abnormal samples with detectable mature neoplastic populations 154(15) Abnormal mature B-cells 154(4) Evaluation of CD5 and CD23 158(5) FCM features suggestive of anti-CD20 therapy 163(1) Abnormal mature T-cells 164(2) Abnormal plasma cells present 166(3) Abnormal blood or bone marrow samples with no detectable neoplastic cells 169(15) Altered cellular composition and abnormal SSC 172(6) Abnormal antigenic maturation in myeloid or erythroid precursors 178(6) Coexisting malignancies 184(3) FCM interpretation and reporting Immature hematopoietic malignancies 187(6) ALL/lymphoblastic lymphoma 188(1) Myeloid malignancies 188(1) AML-M3 189(1) AML with minimal maturation 189(1) AML with maturation 189(1) AML with monocytic differentiation 189(2) AML with erythroid hyperplasia 191(1) AML with megakaryocytic differentiation 192(1) MPD and MDS 193(1) Acute leukemias with a multilineage antigenic profile 193(1) Mature lymphoid malignancies 193(13) B-cell LPD/NHL 195(1) CD10 expression 195(1) Coexpression of CD11c, CD25, and CD103 196(1) Coexpression of CD5 and CD23 196(2) CD5+CD23- B-cell neoplasms 198(2) CD45 and/or pan B-cell antigens markedly downregulated 200(1) Nondescript B-cell phenotype and high FSC 200(1) Nondescript B-cell phenotype and low FSC 200(4) Monoclonal B-cells of undetermined significance 204(1) Plasma cell dyscrasias 204(1) T-cell LPD/NHL 204(1) CD4+ T-cell LPD/NHL 205(1) CD8+ disorders 205(1) CD30+ lymphoma 206(1) FCM reporting 206(5) Suggested Reading 211(4) Appendix 215(4) Index 219