Flow Cytometry in Hematopathology

, by Nguyen, Doyen T.; Diamond, Lawrence W., M.D.; Braylan, Raul C., M.D.

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ISBN: 9781588298553 | 1588298558
Cover: Hardcover
Copyright: 9/26/2007

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The second edition of this volume reflects the recent advances in the FCM analysis of hematopoietic disorders. The chapters have been revised to incorporate new text and figures. The volume is aimed at hematopathologists, hematologists, pathologists, and laboratory technicians.

Preface to the Second Edition	p. ix
Preface to the First Edition	p. xi
Acknowledgments	p. xiii
List of Abbreviations	p. xv
List of Case Studies	p. xvii
Color Plates	p. follow p. 174
Approach to flow cytometry: General considerations
Reasons for the necessity of proper data analysis	p. 2
The pitfalls of the FCM data format of ""percent positive"" per antibody tested	p. 2
General aspects of FCM data analysis and interpretation	p. 4
Other applications of FCM in hematopathology	p. 7
Maturation and differentiation of hematopoietic elements: An overview based on the immunologic markers currently in use in the FCM laboratory	p. 9
FCM immunophenotyping and DNA analysis: Practical aspects that can affect data analysis and interpretation
Sample selection	p. 13
Liquid specimens	p. 14
Solid tissue specimens	p. 15
Preparing nucleated cell suspensions	p. 16
Cell yield and viability	p. 16
Sample staining	p. 16
Surface antigens	p. 17
Intracellular antigens	p. 17
DNA content	p. 18
Data acquisition	p. 19
Calibration	p. 19
Color compensation	p. 19
List mode data collection	p. 20
Exclusion of nonviable cells	p. 21
Antibody panel design	p. 22
Antibody selection	p. 23
Anti-light chain antibodies	p. 23
Fluorochrome conjugation	p. 26
Comprehensive antibody panels	p. 31
Disease-oriented antibody panels	p. 31
Antibody panels oriented by specimen type	p. 32
Tailored panels and add-on testing	p. 34
Minimal residual disease	p. 36
FCM immunophenotyping data representation	p. 37
Analysis panels	p. 37
Color display	p. 38
Approach to DNA data analysis	p. 41
DNA ploidy	p. 42
S-phase	p. 42
FCM data analysis on nearly homogeneous samples
FCM parameters	p. 50
Forward scatter	p. 50
Side scatter	p. 53
Fluorescence P54
Heterogeneous fluorescence intensity (bimodal, variable)	p. 64
Fluorescence dynamic range	p. 71
Strategy to the visual review of FCM immunophenotyping data	p. 71
Common SSC/CD45 patterns	p. 73
Assessment of the blast population	p. 73
Immature neoplastic cells with downregulated CD45	p. 81
SSC/CD45 in mature lymphoid disorders	p. 83
Other dot plot patterns useful in acute leukemia diagnosis	p. 86
Useful antigenic features in AML	p. 86
Myeloid phenotypic abnormalities and MRD detection	p. 95
Precursor B-ALL versus bone marrow B-cell progenitors	p. 98
Useful antigenic features in precursor T-lymphoma/leukemia	p. 99
Evaluation of mature lymphoid malignancies	p. 106
Assessment of surface light chain expression	p. 106
Assessment of pan B-cell antigens	p. 108
Useful antigenic features in mature B-cell malignancies	p. 112
CD10 expression: Follicular center cell lymphomas	p. 112
Pattern of CD20 and CD11c coexpression	p. 117
CD5 expression	p. 123
Aberrant B-cell profile	p. 126
Identification of abnormal mature T-cells	p. 131
Useful antigenic features in mature T-cell malignancies	p. 150
Assessing the biological behavior of mature lymphoid neoplasms	p. 166
Dot plot patterns in histiocytic proliferations and nonhematopoietic malignancies	p. 171
FCM data analysis on heterogeneous specimens
Identifying normal FCM samples	p. 175
Benign/reactive solid lymphoid tissue (e.g., lymph nodes, tonsils)	p. 175
Pattern of CD10/CD20 coexpression: Distinction between FRFH and FCC lymphoma	p. 179
Normal peripheral blood and normal bone marrow	p. 187
Blast region	p. 190
Bone marrow B-cell precursors	p. 192
Lymphocytes	p. 192
Monocytes	p. 208
Plasma cells	p. 209
Erythroid precursors	p. 213
Maturing myeloid cells	p. 216
Abnormal heterogeneous samples with a detectable immature neoplastic population	p. 220
Blasts of lymphoid lineage	p. 221
Blasts of myeloid lineage	p. 222
AML	p. 224
High-grade MDS and MPD with increased blasts	p. 226
Minimal residual disease	p. 233
Abnormal heterogeneous samples with detectable mature neoplastic populations	p. 233
Abnormal mature B-cells	p. 238
Evaluation of CD5 and CD23	p. 238
FCM features suggestive of anti-CD20 therapy	p. 242
Abnormal mature T-cells and NK cells	p. 245
Abnormal plasma cells present	p. 255
Abnormal blood or bone marrow samples with no detectable neoplastic cells	p. 260
Altered cellular composition and abnormal SSC	p. 262
Increased monocytic elements	p. 262
Increased eosinophils	p. 262
Conspicuous basophils or mast cells	p. 265
Abnormal SSC in granulocytes	p. 265
Abnormal antigenic maturation in myeloid or erythroid precursors	p. 273
Antigenic abnormalities in myeloid precursors	p. 273
Antigenic abnormalities in erythroid precursors	p. 280
Identifying low-grade MDS	p. 280
Coexisting malignancies	p. 285
FCM interpretation and reporting
Immature hematopoietic malignancies	p. 289
ALL/lymphoblastic lymphoma	p. 290
Myeloid malignancies	p. 292
AML-M3	p. 292
AML with minimal maturation	p. 292
AML with maturation	p. 292
AML with monocytic differentiation	p. 294
AML with erythroid hyperplasia	p. 295
AML with megakaryocytic differentiation	p. 295
MPD and MDS	p. 295
Acute leukemias with a multilineage antigenic profile	p. 300
Mature lymphoid malignancies	p. 301
B-cell LPD/NHL	p. 302
CD10 expression	p. 302
Coexpression of CD11c, CD25 and CD103	p. 306
Coexpression of CD5 and CD23	p. 306
CD5⁺ CD23^- B-cell neoplasms	p. 307
CD45 and/or pan B-cell antigens markedly downregulated	p. 310
Nondescript B-cell phenotype and high FSC	p. 311
Nondescript B-cell phenotype and low FSC	p. 314
Monoclonal B-cells of undetermined significance	p. 318
Plasma cell dyscrasias	p. 318
T-cell LPD/NHL	p. 320
CD4⁺ T-cell LPD/NHL	p. 321
CD8⁺ disorders	p. 323
CD30⁺ lymphoma	p. 329
FCM reporting	p. 333
Suggested reading	p. 335
Appendix: Using the case studies CD-ROM	p. 341
Index	p. 343
Table of Contents provided by Publisher. All Rights Reserved.

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Flow Cytometry in Hematopathology

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