Practical Flow Cytometry in Haematology Diagnosis

Dr Mike Leach - Department of Haematology, Gartnavel General Hospital, Glasgow, UK

Practical Flow Cytometry in Haematology Diagnosis Dr Mike Leach, Dr Mark Drummond and Dr Allyson Doig Department of Haematology, Gartnavel General Hospital, Glasgow, UK Haematology Department, Beatson West of Scotland Cancer Centre, Glasgow, UK Department of Haematology, Gartnavel Hospital, Glasgow, UK The accurate diagnosis of haematologic malignancies is a complex and challenging task. It routinely involves morphologic, molecular, cytogenetic and flow cytometric expertise. To determine what treatment protocol will be followed, it is vital to integrate, interpret and report these results accurately. Flow cytometry is key in this diagnostic pathway. This book guides the reader as to how flow cytometry results should be interpreted and applied to optimize patient care. At the core of this text is an appreciation of clinical, morphological and immunophenotypic correlation and the importance of constant liaison and discussion between the medical and scientific teams. The authors present a logical and practical approach to the diagnosis of blood disorders (both neoplastic and reactive) and evaluate the diagnostic applications of flow cytometry. Practical Flow Cytometry in Haematology Diagnosis provides: ? A clinical reference source on all aspects of flow cytometry, covering both malignant and benign conditions ? Carefully chosen real-life cases in each chapter, complemented by high quality morphological images ? Help in making a diagnosis, together with an understanding of the limitations of the technique and the potential pitfalls All those who instigate, perform, interpret or act upon flow cytometry patient material will find this book an invaluable guide. Titles of related interest Ortolani: Flow Cytometry of Hematological Malignancies (2011) ISBN: 978-1-4443-3588-0 Kottke-Marchant: Laboratory Hematology Practice (2012) ISBN: 978-1-4051-6218-0

Preface, vii
 
Acknowledgements, viii
 
1 Introduction, 1
 
References, 2
 
2 Principles of Flow Cytometry, 3
 
Introduction, 3
 
Sample preparation, 3
 
The flow cytometer, 4
 
The fluidic system, 4
 
The optical system, 5
 
The electronic system, 9
 
Data display, 9
 
Acknowledgements, 18
 
References, 19
 
3 Limitations, 20
 
Introduction, 20
 
Clinical context issues, 20
 
Sampling issues, 20
 
Technical issues, 25
 
Interpretation issues, 28
 
Conclusion, 29
 
References, 29
 
4 Normal Blood and Bone Marrow Populations, 31
 
Normal stem and precursor cell populations, 31
 
References, 41
 
5 Acute Leukaemia, 43
 
Introduction, 43
 
Identification of neoplastic precursor cells, 45
 
Lineage assignment of acute leukaemia, 52
 
Acute myeloid leukaemia, 54
 
Precursor lymphoid neoplasms (acute lymphoblastic leukaemia/lymphoma), 77
 
Examination of cerebro-spinal fluid and serous effusions, 88
 
Identification of genetic abnormalities by flow, 92
 
Conclusion, 93
 
References, 93
 
6 Chronic Lymphoid Leukaemias and Exfoliating Lymphoma, 100
 
Normal peripheral blood lymphoid populations, 100
 
Identification of clonal lymphoid populations, 101
 
Identification of clonal B-cell disorders, 101
 
CD5 positive B-cell lymphoproliferative disorders, 102
 
CD10 positive B-cell lymphoproliferative disorders, 116
 
B-cell disorders identified using an extended B-cell panel, 121
 
Identification of clonal T-cell disorders, 125
 
CD4 positive T-cell disorders, 128
 
CD8 positive T-cell disorders, 136
 
CD4/CD8 positive T-cell disorders, 141
 
CD4/8 negative disorders, 141
 
Flow cytometric assessment of serous effusions, 146
 
Flow cytometric assessment of cerebrospinal fluid, 146
 
Summary, 149
 
References, 149
 
7 Myelodysplastic Syndromes and Myeloproliferative Neoplasms, 152
 
Introduction, 152
 

Myelodysplastic syndromes, 152
 
Myelodysplastic/myeloproliferative neoplasms, 163
 
Chronic myeloid leukaemia, 165
 
The BCR-ABL negative myeloproliferative neoplasms, 166
 
Mastocytosis, 167
 
Summary, 170
 
References, 170
 
8 Disorders of Plasma Cells, 173
 
Plasma cell disorders, 173
 
Current diagnostic criteria, 173
 
Plasma cell morphology, 175
 
The applications of immunophenotyping to plasma cell disorders, 178
 
References, 183
 
9 Minimal Residual Disease, 184
 
Introduction, 184
 

Minimal residual disease analysis in the management of acute lymphoblastic leukaemia, 187
 
Conclusions, 199
 
Acknowledgements, 200
 
References, 200
 
10 Red Cells, Leucocytes and Platelets, 202
 
Paroxysmal nocturnal haemoglobinuria, 202
 
Red cell membrane disorders, 208
 
Foetal maternal haemorrhage, 211
 
Lymphocyte subset analysis
 
and immunodeficiency, 214
 
Haemopoietic stem cell enumeration, 214
 
Granulocyte disorders, 215
 
Platelet disorders, 219
 
References, 221
 
11 Reactive and Non-neoplastic Phenomena, 225
 
Peripheral blood, 225
 
Bone marrow, 231
 
Pleural and pericardial fluids, 236
 
Bronchoalveolar lavage specimens, 238
 
Cerebrospinal fluid, 238
 
References, 240
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