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Practical Flow Cytometry in Haematology Diagnosis



Practical Flow Cytometry in Haematology Diagnosis

Mike Leach, Mark Drummond, Allyson Doig

ISBN: 978-1-118-48799-0 January 2013 Wiley-Blackwell 256 Pages

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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.

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

Index, 243

“I would particularly recommend this book to haematology trainees, as an introduction to, but not as a replacement for, a period spent in an immunophenotyping laboratory.”  (British Journal of Haematology, 14 November 2014)

“There is no doubt that this book will find its way firmly onto the bookshelf in every laboratory, and into the work bags of many trainees.”  (Haem Trainee, 1 January 2014)

“This is a concise and very practical guide to the use of flow cytometry in hematological and hematopathological diagnosis. While not as detailed in immunophenotyping intricacies as other books of its kind, it shines with its practical approach and expert guidance applied to current clinical practice.”  (Doody’s, 19 July 2013)