Practical Flow Cytometry in Haematology E-Book

Mike Leach

• Herausgeber: John Wiley & Sons
• Kategorie: Fachliteratur
• Sprache: Englisch

Seitenzahl: 508

Veröffentlichungsjahr: 2015

Beschreibung

The analysis of blood, bone marrow and tissue fluid specimens requires a multi-faceted approach with the integration of scientific data from a number of disciplines. No single discipline can operate in isolation or errors will occur. Flow cytometry is in a privileged position in that it can provide rapid analysis of specimens and it is often the first definitive investigation to produce results and help formulate a working diagnosis.

This companion text to Practical Flow Cytometry in Haematology Diagnosis contains 100 worked examples drawn from real clinical cases presenting to the authors’ institution. Cases are illustrated with peripheral blood and bone marrow cytology, tissue pathology and cytogenetic and molecular data, which are integrated to generate, where appropriate, a diagnosis based on the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. The spectrum of clinical cases includes adult and paediatric patients, and both neoplastic and reactive disorders. The cases appear in no particular order to challenge the reader to make their own diagnosis.

The reader will review May−Grünwald−Giemsa (MGG)-stained films of peripheral blood and bone marrow aspirates presented alongside flow cytometric data and haematoxylin and eosin (H&E)-stained bone marrow and other tissue biopsy sections. Immunohistochemistry is used to further clarify the tissue lineage and cell differentiation. Cytogenetic studies using metaphase preparations are used to identify translocations and chromosome gains and losses whilst interphase fluorescence in situ hybridisation (FISH) studies and polymerase chain reaction (PCR) are used to identify gene fusions, gene rearrangements and deletions. Each case concludes with a discussion of the features that are important to making a diagnosis. The cases are also listed according to disease classification in the appendix so that the text can also be used as a reference.

Practical Flow Cytometry in Haematology: 100 Worked Examples:

• Provides a practical, example-based resource for flow cytometry
• Demonstrates how flow cytometry results should be interpreted and applied to optimize patient care
• Includes both malignant and benign conditions
• Can be used in conjunction with Practical Flow Cytometry in Haematology Diagnosis, by the same author team (ISBN 9780470671207)

Practical Flow Cytometry in Haematology: 100 Worked Examples is ideal for practicing haematologists and histopathologists with an interest in haematopathology, but particularly directed at trainee haematologists and scientists preparing for FRCPath and related examinations.

Leseprobe

Table of Contents

Title Page

Copyright

Preface

Acknowledgement

List of Abbreviations

Technical Notes

Full blood counts

Biochemistry and immunology studies

Flow cytometry analysis

Immunohistochemistry in paraffin-embedded formalin fixed tissue

Laboratory Values

Case 1

Laboratory investigations

Imaging

Flow cytometry

Histopathology

FISH studies

Discussion

Final diagnosis

Case 2

Laboratory results

Imaging

Pleural fluid biochemistry and cytology

Morphology (pleural fluid)

Flow cytometry (pleural fluid)

Lymph node biopsy

Discussion

Final diagnosis

Case 3

Laboratory data

Marrow aspirate

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis

Bone marrow trephine biopsy

Discussion

Final diagnosis

References

Case 4

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Discussion

Final diagnosis

Case 5

Laboratory data

Imaging

Blood film

Flow cytometry (peripheral blood)

Lymph node core histology

Discussion

Final diagnosis

Case 6

Laboratory results

Flow cytometry (bone marrow aspirate)

Peripheral blood morphology

Flow cytometry (peripheral blood)

Histopathology

Discussion

Final diagnosis

Case 7

Laboratory data

Blood film

Platelet aggregation studies

Flow cytometry studies

Molecular studies

Management

Discussion

Final diagnosis

Reference

Case 8

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry on peripheral blood

Cytogenetic analysis on peripheral blood

Clinical course

Discussion

Final diagnosis

Case 9

Laboratory data

Blood film

Flow cytometry

Bone marrow trephine biopsy

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

References

Case 10

Laboratory investigations

Blood film

Flow cytometry (peripheral blood)

Imaging

Lymph node core biopsy

Bone marrow aspirate and trephine biopsy

Molecular studies

Classical cytogenetics

Clinical course

Discussion

Final diagnosis

References

Case 11

Laboratory results

Blood film

Flow cytometry (peripheral blood)

FISH

Imaging

Histopathology

Discussion

Final diagnosis

References

Case 12

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 13

Laboratory investigations

Blood film

Bone marrow aspirate

Flow cytometry

Histopathology

Discussion

Final diagnosis

Case 14

Laboratory results

Blood film

Flow cytometry

Imaging

Clinical course

Bone marrow aspirate and trephine biopsy

Cervical lymph node biopsy

Discussion

Final diagnosis

Case 15

Laboratory data

Imaging

Blood film

Bone marrow aspirate

Flow cytometry (peripheral blood and marrow)

Histopathology

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

Reference

Case 16

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Discussion

Final diagnosis

Case 17

Laboratory results

Blood film

Imaging

Flow cytometry (peripheral blood)

Discussion

Final diagnosis

Reference

Case 18

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 19

Laboratory data

Imaging

Bone marrow aspirate

Flow cytometry

Discussion

Final diagnosis

Reference

Case 20

Laboratory results

Blood film

Flow cytometry (peripheral blood)

Imaging

Bone marrow trephine biopsy

Discussion

Final diagnosis

Reference

Case 21

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Discussion

Final diagnosis

References

Case 22

Laboratory data

Blood film/bone marrow aspirate

Flow cytometry

Genetic studies

Discussion

Final diagnosis

References

Case 23

Laboratory results

Imaging

Flow cytometry (pleural fluid)

Discussion

Final diagnosis

Case 24

Laboratory data

Blood film

Imaging

Morphology (bone marrow aspirate)

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis

Discussion

Final diagnosis

Reference

Case 25

Laboratory data

Imaging

Blood film

Marrow aspirate

Bone marrow trephine biopsy

Discussion

Diagnosis

References

Case 26

Laboratory investigations

Imaging

Flow cytometry

Bone marrow trephine biopsy

FISH studies

Discussion

Final diagnosis

References

Case 27

Laboratory data

Blood film

Flow cytometry

Differential diagnosis

Molecular studies

Discussion

Final diagnosis

Reference

Case 28

Laboratory results

Imaging

Bone marrow aspirate

Flow cytometry (bone marrow)

Bone marrow trephine biopsy

Core biopsy of retroperitoneal tissue

Discussion

Final diagnosis

Case 29

Basic laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (blood)

Flow cytometry (bone marrow aspirate)

Cytogenetics

FISH studies

Discussion

Final diagnosis

Reference

Case 30

Laboratory data

Blood film

Differential diagnosis

Flow cytometry

Virology

Discussion

Final diagnosis

Case 31

Laboratory results

Imaging

Pleural fluid morphology

Pleural fluid flow cytometry

Immunocytochemistry and immunohistochemistry (pleural fluid)

Cytogenetic analysis (pleural fluid)

Discussion

Final diagnosis

Reference

Case 32

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Histopathology

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 33

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (peripheral blood and bone marrow)

Imaging

Histopathology

FISH studies

Cytogenetics

Discussion

Final diagnosis

Case 34

Laboratory results

Imaging

Blood film

Flow cytometry (peripheral blood)

Flow cytometry (bone marrow aspirate)

Bone marrow trephine biopsy

Clinical course

Histopathology

Discussion

Final diagnoses

Reference

Case 35

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Imaging

Bone marrow trephine biopsies (bilateral)

Discussion

Final diagnosis

Case 36

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Histopathology

Cytogenetics

FISH studies

Molecular studies

Discussion

Final diagnosis

References

Case 37

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Cytogenetics

Additional investigations

Discussion

Final diagnosis

References

Case 38

Laboratory results

Blood film

Flow cytometry

Molecular studies

Discussion

Final diagnosis

References

Case 39

Laboratory results

Blood film

Imaging

Flow cytometry (peripheral blood)

Bone marrow trephine biopsy

Histopathology

Discussion

Final diagnosis

Reference

Case 40

Laboratory results

Blood film

Flow cytometry (peripheral blood)

Histopathology

Imaging

Discussion

Final diagnosis

Case 41

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (peripheral blood)

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

Reference

Case 42

Laboratory data

Bone marrow aspirate

Trephine biopsy

Flow cytometry

Discussion

Final diagnosis

References

Case 43

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Histopathology

Cytogenetic analysis

Discussion

Final diagnosis

Reference

Case 44

Laboratory investigations

Blood film

Flow cytometry

Imaging

Histopathology

Discussion

Final diagnosis

Reference

Case 45

Laboratory data

Bone marrow aspirate

Flow cytometry on marrow aspirate at diagnosis

Cytogenetic profile at diagnosis

Flow cytometry on marrow aspirate at day 100

Discussion

Final diagnosis

Case 46

Laboratory results

Blood film

Flow cytometry (peripheral blood)

Imaging

Bone marrow trephine biopsy

Discussion

Final diagnosis

Reference

Case 47

Laboratory data

Imaging

CSF cytospin

Flow cytometry

Genetic studies

Discussion

Final diagnosis

References

Case 48

Laboratory data

Bone marrow aspirate

Flow cytometry

Histopathology

Discussion

Final diagnosis

References

Case 49

Laboratory data

Blood film

Imaging

Flow cytometry (peripheral blood and bone marrow)

Cytogenetics

Molecular

Discussion

Final diagnosis

Reference

Case 50

Initial laboratory data

Subsequent course

Repeat laboratory data

Blood film

Flow cytometry

Discussion

Final diagnosis

Reference

Case 51

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (peripheral blood)

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 52

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Bone marrow aspirate

Flow cytometry (bone marrow)

Bone marrow trephine biopsy

Skeletal survey/MRI

Discussion

Final diagnoses

Case 53

Laboratory results

Blood film

Flow cytometry

Imaging

Bone marrow aspirate and trephine biopsy

Discussion

Final diagnosis

References

Case 54

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Marrow trephine biopsy

Cytogenetics

Discussion

Final diagnosis

Reference

Case 55

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Molecular studies

Imaging

Bone marrow aspirate

Discussion

Final diagnosis

Reference

Case 56

Laboratory results

Imaging

Histopathology

Clinical course

Flow cytometry

Discussion

Final diagnosis

Case 57

Clinical images

Blood film

Histopathology

Bone marrow aspirate

Flow cytometry

Discussion

Final diagnosis

References

Case 58

Laboratory results

Peripheral blood morphology

Bone marrow trephine biopsy

Discussion

Final diagnosis

Case 59

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Discussion

Final diagnosis

Case 60

Laboratory data

Blood film

Flow cytometry

Final diagnosis (main case)

Case 61

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis (marrow aspirate)

Molecular analysis

Discussion

Final diagnosis

References

Case 62

Laboratory data

Blood film

Bone marrow aspirate and trephine biopsy

Bone marrow aspirate

Flow cytometry

Bone marrow trephine biopsy

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 63

Laboratory data

Blood film

Platelet aggregation studies

Flow cytometry studies

Discussion

Final diagnosis

Case 64

Laboratory data

Bone marrow aspirate at diagnosis

Histopathology at diagnosis

Bone marrow aspirate following induction chemotherapy

Flow cytometry at remission

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

Case 65

Laboratory data

Blood film

Imaging

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Bone marrow trephine biopsy

Discussion

Final diagnosis

Reference

Case 66

Laboratory investigations

Imaging

Flow cytometry

Histopathology

Molecular studies

Discussion

Final diagnosis

References

Case 67

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (marrow aspirate)

Cytogenetic analysis

Discussion

Final diagnosis

Case 68

Laboratory investigations

Flow cytometry

Bone marrow trephine biopsy

Cytogenetic analysis

Discussion

Final diagnosis

Case 69

Laboratory data

Histopathology

Bone marrow aspirate

Flow cytometry

Discussion

Final diagnosis

Reference

Case 70

Laboratory data

CSF cytospin morphology

Flow cytometry

Bone marrow aspirate

Skin lesions

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 71

Laboratory data

Blood film

Bone marrow aspirate and trephine biopsy

Flow cytometry (peripheral blood)

Urine haemosiderin

Discussion

Final diagnoses

Case 72

Laboratory data

Blood film

Imaging

Bone marrow aspirate

Flow cytometry (peripheral blood and bone marrow)

Histopathology

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 73

Laboratory data

Blood film

Flow cytometry (pleural fluid)

Cytology

Clinical course

Discussion

Final diagnosis

Reference

Case 74

Laboratory data

Blood film

Imaging

Bone marrow aspirate

Flow cytometry

Histopathology

Cytogenetic analysis (marrow aspirate)

Discussion

Final diagnosis

References

Case 75

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Discussion

Final diagnosis

Case 76

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry on blood and bone marrow aspirate

Cytogenetic analysis

Immediate clinical course

CSF cytospin

Flow cytometry (CSF)

Bone marrow aspirate (at CNS relapse)

Discussion

Final diagnosis

References

Case 77

Laboratory results

Imaging

Pleural fluid aspirate

Flow cytometry (pleural fluid)

Histopathology

Discussion

Final diagnosis

References

Case 78

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (bone marrow)

Histopathology

Cytogenetic analysis

Discussion

Final diagnosis

Reference

Case 79

Laboratory results

Imaging

CSF analysis

Flow cytometry (cerebrospinal fluid)

Discussion

Final diagnosis

Case 80

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (marrow aspirate)

Bone marrow trephine biopsy

Cytogenetic analysis

Molecular studies

Additional investigations

Discussion

Final diagnosis

Reference

Case 81

Laboratory investigations

Imaging

Flow cytometry

Discussion

Final diagnosis

Case 82

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (peripheral blood)

Trephine biopsy

Cytogenetic analysis

Discussion

Final diagnosis

References

Case 83

Laboratory data

Blood film

Imaging

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Trephine biopsy

Lymph node biopsy

Discussion

Final diagnosis

Case 84

Laboratory results

Imaging

Peripheral blood morphology

Flow cytometry (peripheral blood)

Histopathology

Molecular studies

Discussion

Final diagnosis

References

Case 85

Laboratory data

Blood film

Imaging

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis and FISH

Discussion

Final diagnosis

Reference

Case 86

Laboratory results

Blood film

Flow cytometry (peripheral blood)

Histopathology

Imaging

Discussion

Final diagnosis

Case 87

Laboratory data

Imaging

Blood film

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

References

Case 88

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Additional investigations

Discussion

Final diagnosis

Case 89

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry

Histopathology

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

References

Case 90

Laboratory data

Blood film

Flow cytometry

Bone marrow aspirate and trephine biopsy

Imaging

Histopathology

Discussion

Final diagnosis

References

Case 91

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry at diagnosis

Cytogenetic analysis

Molecular studies

Treatment course

Discussion

Final diagnosis

Case 92

Laboratory results

Blood and bone marrow examination

Imaging

Clinical course

Flow cytometry (peripheral blood)

Bone marrow trephine biopsy

Discussion

Final diagnosis

References

Case 93

Laboratory data

Blood film

Imaging

Histopathology

Cytogenetic analysis

Bone marrow aspirate

Flow cytometry (bone marrow aspirate)

Trephine biopsies

Discussion

Final diagnosis

References

Case 94

Laboratory data

Blood film

Bone marrow aspirate

Flow cytometry (marrow aspirate)

Histopathology

Cytogenetics

Discussion

Final diagnosis

References

Case 95

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Bone marrow aspirate

Cytogenetic analysis

Molecular studies

Discussion

Final diagnosis

References

Case 96

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Histopathology

Discussion

Final diagnosis

Case 97

Laboratory data

Blood film

Imaging

Marrow aspirate

Flow cytometry (bone marrow aspirate)

Bone marrow trephine biopsy

Cytogenetic analysis

Discussion

Final diagnosis

Case 98

Laboratory investigations

Blood film

Imaging

Bone marrow aspirate

Flow cytometry

Histopathology

FISH and cytogenetics

Discussion

Final diagnosis

Reference

Case 99

Laboratory data

Blood film

CT imaging

Flow cytometry (pleural fluid)

Bone marrow aspirate and trephine biopsy

Omental biopsy

Cytogenetic analysis

FISH studies

Discussion

Reference

Final diagnosis

Case 100

Laboratory data

Blood film

Flow cytometry (peripheral blood)

Bone marrow aspirate

Histopathology

Cytogenetics

FISH studies

Discussion

Final diagnosis

Antibodies Used in Immunohistochemistry Studies

Flow Cytometry Antibodies

Molecular Terminology

Classification of Cases According to Diagnosis

Myeloproliferative neoplasms

Myeloid and lymphoid neoplasms with eosinophilia and abnormalities of PDGRFA, PDGRFB or FGFR1

Myelodysplastic/myeloproliferative neoplasms

Myelodysplastic syndromes

Acute myeloid leukaemia and related precursor neoplasms

Acute leukaemias of ambiguous lineage

Precursorlymphoid neoplasms

Mature B-cell neoplasms

Plasma cell neoplasms

Mature T- and NK-cell neoplasms

Hodgkin lymphoma

Immunodeficiency-associated lymphoproliferative disorders

Platelet disorders

Red cell disorders

Marrow aplasia

Non-haemopoietic tumours

Reactive phenomena

Index

End User License Agreement

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Guide

Cover

Table of Contents

Preface

Begin Reading

List of Illustrations

Technical Notes

Figure 1.1 Visual representation of strength of fluorescence in flow cytometry (not actual patient specimens), showing an isotype control and eight CD19-positive samples which show fluorescence intensity with CD20 varying from negative to bright. (a) Isotype control, used to set thresholds. (b) Negative (consistent with a CD19-positive, CD20-negative B-cell precursor neoplasm). (c) Partial positive, indicating that CD20 antigen expression varies from negative to positive (consistent with a precursor B-cell neoplasm). (c adjusted) Indicating that the threshold for positivity might be reduced by the cytometrist where a discrete dim positive population is identified. (d) Dim CD20 antigen expression (consistent with chronic lymphocytic leukaemia). (e) Moderate intensity, indicating medium strength of CD20 antigen expression (consistent with B-cell non-Hodgkin lymphoma). (f) bright, indicating strong CD20 antigen expression (consistent with hairy cell leukaemia). (g) Two distinct populations, one partial and dim and one bright (could indicate two unrelated B-lineage neoplasms or transformation of a low grade lymphoma). (h) Contrasting with (g), a heterogeneous single population with fluorescence intensity varying from negative to moderate with a minority being bright.

Case 1

Figure 1.1 CXR.

Figure 1.2 CT.

Figure 1.3 MGG, ×500.

Figure 1.4 MGG, ×500.

Figure 1.5 MGG, ×500.

Figure 1.6 MGG, ×500.

Figure 1.7 FSC/SSC.

Figure 1.8 CD3/CD45.

Figure 1.9 CD2/CD19.

Figure 1.10 CD7/CD16.

Figure 1.11 H&E, ×400.

Figure 1.12 CD2, ×400.

Figure 1.13 CD30, ×400.

Figure 1.14 ALK, ×400.

Figure 1.15 CD30, ×100.

Case 2

Figure 2.1 CT.

Figure 2.2 MGG, ×500.

Figure 2.3 MGG, ×500.

Figure 2.4 MGG, ×500.

Figure 2.5 MGG, ×500.

Figure 2.6 FSC/SSC.

Figure 2.7 CD19/SSC.

Figure 2.8 CD10/HLA-DR.

Figure 2.9 Kappa/lambda.

Figure 2.10 Serous (left) and chylous (right) pleural effusions.

Figure 2.11 MGG, ×500.

Figure 2.12 MGG, ×500.

Case 3

Figure 3.1 MGG, ×1000.

Figure 3.2 MGG, ×1000.

Figure 3.3 MGG, ×1000.

Figure 3.4 MGG, ×1000.

Figure 3.5 H&E, ×40.

Figure 3.6 CD34, ×200.

Figure 3.7 MPO, ×200.

Figure 3.8 CD15, ×200.

Figure 3.9 CD68, ×200.

Case 4

Figure 4.1 MGG, x1000.

Figure 4.2 MGG, x1000.

Figure 4.3 MGG, x1000.

Figure 4.4 MGG, x1000.

Figure 4.5 CD138/SSC.

Figure 4.6 CD19/CD45.

Figure 4.7 CD38/CD56.

Case 5

Figure 5.1 CT.

Figure 5.2 MGG, x1000.

Figure 5.3 MGG, x1000.

Figure 5.4 MGG, x1000.

Figure 5.5 MGG, x1000.

Figure 5.6 H&E, x25.

Figure 5.7 H&E, x400.

Figure 5.8 H&E, x400.

Figure 5.9 EBER ISH, x40.

Case 6

Figure 6.1 FSC/SSC.

Figure 6.2 MGG, ×1000.

Figure 6.3 MGG, ×1000.

Figure 6.4 MGG, ×1000.

Figure 6.5 Blood, FSC/SSC.

Figure 6.6 H&E, ×400.

Figure 6.7 Reticulin, ×400.

Case 7

Figure 7.1 MGG, ×500.

Figure 7.2 MGG, ×1000.

Figure 7.3 Platelet aggregation.

Case 8

Figure 8.1 MGG, ×1000.

Figure 8.2 MGG, ×1000.

Figure 8.3 H&E, ×40.

Figure 8.4 H&E, ×200.

Figure 8.5 CD34, ×200.

Figure 8.6 MPO, ×200.

Figure 8.7 CD33, ×200.

Figure 8.8 CD15, ×200.

Case 9

Figure 9.1 MGG, ×1000.

Figure 9.2 MGG, ×1000.

Figure 9.3 MGG, ×1000.

Figure 9.4 MGG, ×1000.

Figure 9.5 H&E, ×100.

Figure 9.6 H&E, ×100.

Figure 9.6 H&E, ×100.

Figure 9.7 H&E, ×200.

Figure 9.8 H&E, ×400.

Figure 9.9 H&E, ×100.

Case 10

Figure 10.1 MGG, ×1000.

Figure 10.2 MGG, ×1000.

Figure 10.3 MGG, ×1000.

Figure 10.4 H&E, ×500.

Figure 10.5 H&E, ×500.

Figure 10.6 Abbott Molecular Vysis LSI IGH/BCL2 Dual-Color, Dual-Fusion Translocation Probe.

Figure 10.7 Metaphase cytogenetic preparation showing a three-way translocation, t(8;14;18)(q24;q32;q21).

Figure 10.8 MGG, ×1000.

Case 11

Figure 11.1 MGG, ×500.

Figure 11.2 MGG, ×500.

Figure 11.3 MGG, ×1000.

Figure 11.4 MGG, ×1000.

Figure 11.5 H&E, ×40.

Figure 11.6 H&E, ×200.

Figure 11.7 H&E, ×400.

Figure 11.8 Cyclin D1, ×200.

Case 12

Figure 12.1 MGG, ×1000.

Figure 12.2 MGG, ×1000.

Figure 12.3 MGG, ×1000.

Figure 12.4 MGG, ×1000.

Case 13

Figure 13.1 MGG, ×500.

Figure 13.2 MGG, ×1000.

Figure 13.3 MGG, ×1000.

Figure 13.4 MGG, ×1000.

Figure 13.5 CD45/SSC.

Figure 13.6 CXR.

Figure 13.7 H&E, ×100.

Figure 13.8 H&E, ×500.

Figure 13.9 CD56, ×400.

Figure 13.10 CAM5.2, ×400.

Case 14

Figure 14.1 MGG, ×1000.

Figure 14.2 MGG, ×1000.

Figure 14.3 H&E, ×40.

Figure 14.4 H&E, ×400.

Figure 14.5 CD20, ×400.

Figure 14.6 CD23, ×400.

Figure 14.7 H&E, ×20.

Figure 14.8 CD20, ×40.

Figure 14.9 CD21, ×40.

Figure 14.10 Ki-67, ×40.

Case 15

Figure 15.1 MGG, ×1000.

Figure 15.2 MGG, ×1000.

Figure 15.3 MGG, ×1000.

Figure 15.4 MGG, ×1000.

Figure 15.5 MGG, ×500.

Figure 15.6 MGG, ×1000.

Figure 15.7 MGG, ×1000.

Figure 15.8 FSC/SSC.

Figure 15.9 CD45/SSC.

Figure 15.10 H&E, ×400.

Figure 15.11 H&E, ×400.

Case 16

Figure 16.1 MGG, ×500.

Figure 16.2 MGG, ×500.

Figure 16.3 MGG, ×1000.

Figure 16.4 MGG, ×500.

Figure 16.5 MGG, ×1000.

Figure 16.6 MGG, ×1000.

Case 17

Figure 17.1 MGG, ×500.

Figure 17.2 MGG, ×500.

Figure 17.3 MGG, ×1000.

Figure 17.4 MGG, ×1000.

Figure 17.5 FSC/SSC.

Figure 17.6 CD4/CD26.

Figure 17.7 CD7/CD16.

Case 18

Figure 18.1 MGG, ×1000.

Figure 18.2 MGG, ×1000.

Figure 18.3 MGG, ×1000.

Figure 18.4 MGG, ×1000.

Figure 18.5 MGG, ×1000.

Figure 18.6 CD34/SSC.

Figure 18.7 CD117/CD15.

Case 19

Figure 19.1 CD138/SSC.

Figure 19.2 CD19/CD45.

Figure 19.3 CD56/CD38.

Case 20

Figure 20.1 MGG, ×500.

Figure 20.2 MGG, ×1000.

Figure 20.3 MGG, ×1000.

Figure 20.4 MGG, ×1000.

Figure 20.5 H&E, ×200.

Figure 20.6 CD20, ×200.

Figure 20.7 CD138, ×200.

Figure 20.8 Lambda, ×200.

Case 21

Figure 21.1 MGG, ×1000.

Figure 21.2 MGG, ×1000.

Figure 21.3 MGG, ×1000.

Figure 21.4 MGG, ×1000.

Case 22

Figure 22.1 MGG, ×1000.

Figure 22.2 MGG, ×1000.

Figure 22.3 MGG, ×1000.

Figure 22.4 MGG, ×1000.

Figure 22.5 MGG, ×1000.

Figure 22.6 FISH,

PML

(15q24.1)

/RARA

(17q21.1-17q21.2) translocation dual fusion probes. This cell shows a single red and a single green signal representing the normal chromosomes 15 and 17, respectively. The arrows indicate the two fusion signals representing the reciprocal translocations.

Figure 22.7 Metaphase cytogenetics.

Case 23

Figure 23.1 Contrast CT.

Figure 23.2 PET/CT.

Figure 23.3

Case 24

Figure 24.1 CT bone windows.

Figure 24.2 MRI.

Figure 24.3 MGG, ×500.

Figure 24.4 MGG, ×500.

Figure 24.5 MGG, ×1000.

Figure 24.6 MGG, x1000.

Figure 24.7 CD138/SSC.

Figure 24.8 FSC/SSC.

Figure 24.9 CD45/CD19.

Figure 24.10 CD38/CD56.

Figure 24.11 H&E, ×40.

Figure 24.12 H&E, ×400.

Figure 24.13 CD138, ×400.

Figure 24.14 CD56, ×400.

Figure 24.15 Lambda, ×400.

Figure 24.16 Ki-67, ×400.

Case 25

Figure 25.1 CT (bone windows).

Figure 25.2 MGG, ×100.

Figure 25.3 H&E, ×500.

Figure 25.4 H&E, ×500.

Figure 25.5 CD117, ×200.

Figure 25.6 Mast cell tryptase, ×200.

Case 26

Figure 26.1 MGG, ×1000.

Figure 26.2 MGG, ×1000.

Figure 26.3 CT.

Figure 26.4 H&E, ×400.

Figure 26.5 CD20, ×400.

Figure 26.6 CD5, ×400.

Figure 26.7 Cyclin D1, ×400.

Case 27

Figure 27.1 MGG, ×500.

Figure 27.2 MGG, ×500.

Case 28

Figure 28.1 MGG, ×500.

Figure 28.2 MGG, ×1000.

Figure 28.3 MGG, ×1000.

Figure 28.4 CD10/HLA-DR.

Figure 28.5 Kappa/lambda.

Figure 28.6 H&E, ×40.

Figure 28.7 H&E, ×400.

Figure 28.8 BCL2, ×200.

Figure 28.9 BCL6, ×200.

Figure 28.10 Ki-67, ×100.

Figure 28.11 H&E, ×40.

Figure 28.12 H&E, ×100.

Figure 28.13 H&E, ×400.

Figure 28.14 CD21, ×40.

Figure 28.15 Ki-67, ×40.

Case 29

Figure 29.1 MGG, ×1000.

Figure 29.2 MGG, ×1000.

Figure 29.3 MGG, ×1000.

Figure 29.4 MGG, ×1000.

Figure 29.5 MGG, ×1000.

Figure 29.6 MGG, ×1000.

Figure 29.7 MGG, ×1000.

Figure 29.8 MGG, ×1000.

Figure 29.9 CD14/CD64.

Figure 29.10 CD38/CD56.

Figure 29.11 CD14/CD64.

Case 30

Figure 30.1 MGG, ×500.

Figure 30.2 MGG, ×500.

Figure 30.3 MGG, ×1000.

Figure 30.4 MGG, ×1000.

Case 31

Figure 31.1 MGG, ×1000.

Figure 31.2 MGG, ×1000.

Figure 31.3 Papanicolaou, ×400.

Figure 31.4 Giemsa, ×500.

Figure 31.5 CD20, ×400.

Figure 31.6 BCL2, ×400.

Figure 31.7 BCL6, ×400.

Figure 31.8 Ki-67, ×400.

Case 32

Figure 32.1 MGG, ×1000.

Figure 32.2 MGG, ×1000.

Figure 32.3 MGG, ×1000.

Figure 32.4 MGG, ×1000.

Figure 32.5 MGG, ×1000.

Figure 32.6 MGG, ×1000.

Figure 32.7 MGG, ×1000.

Figure 32.8 MGG, ×1000.

Figure 32.9 MGG, ×1000.

Figure 32.10 MGG, ×1000.

Case 33

Figure 33.1 MGG, ×500.

Figure 33.2 MGG, ×500.

Figure 33.3 MGG, ×1000.

Figure 33.4 MGG, ×1000.

Figure 33.5 Abbott Vysis

MYC

break-apart probe.

Figure 33.6 Abbott Vysis

IGH/MYC

CEP 8 dual-fusion probe.

Figure 33.7 Metaphase cytogenetic preparation showing t(8;14)(q24;q32).

Case 34

Figure 34.1 MGG, ×500.

Figure 34.2 MGG, ×500.

Figure 34.3 MGG, ×1000.

Figure 34.4 MGG, ×1000.

Figure 34.5 H&E, ×40.

Figure 34.6 H&E, ×200.

Figure 34.7 H&E, ×400.

Figure 34.8 CD3, ×100.

Figure 34.9 CD7, ×100.

Figure 34.10 Cyclin D1, ×100.

Figure 34.11 PD-1 (CD279), ×400.

Figure 34.12 CD20, ×40.

Figure 34.13 CD5, ×400.

Figure 34.14 Cyclin D1, ×400.

Case 35

Figure 35.1 MGG, ×500.

Figure 35.2 MGG, ×1000.

Figure 35.3 CD45/SSC.

Figure 35.4 CD56/CD38.

Figure 35.5 CD117/CD15.

Figure 35.6 MRI.

Figure 35.7 CT.

Figure 35.8 MRI.

Figure 35.9 H&E, ×100.

Figure 35.10 H&E, ×500.

Figure 35.11 NB84, ×200.

Figure 35.12 CD56, ×200.

Case 36

Figure 36.1 MGG, ×1000.

Figure 36.2 MGG, ×1000.

Figure 36.3 MGG, ×1000.

Figure 36.4 MGG, ×1000.

Figure 36.5 MGG, ×1000.

Figure 36.6 MGG, ×1000.

Case 37

Figure 37.1 MGG, ×1000.

Figure 37.2 MGG, ×1000.

Figure 37.3 MGG, ×1000.

Figure 37.4 MGG, ×1000.

Figure 37.5 MGG, ×1000.

Figure 37.6 MGG, ×1000.

Case 38

Figure 38.1 MGG, ×1000.

Figure 38.2 MGG, ×1000.

Figure 38.3 MGG, ×1000.

Figure 38.4 CD4/CD8.

Figure 38.5 CD2/CD26.

Figure 38.6 H&E, ×40.

Figure 38.7 H&E, ×400.

Figure 38.8 CD3, ×400.

Figure 38.9 CD57, ×400.

Case 39

Figure 39.1 MGG, ×1000.

Figure 39.2 MGG, ×1000.

Figure 39.3 CT.

Figure 39.4 FSC/SSC.

Figure 39.5 CD5/CD20.

Figure 39.6 H&E, ×100.

Figure 39.7 CD20, ×100.

Figure 39.8 H&E, ×25.

Figure 39.9 H&E, ×100.

Figure 39.10 H&E, ×400.

Figure 39.11 CD20, ×400.

Case 40

Figure 40.1 MGG, ×1000.

Figure 40.2 MGG, ×1000.

Figure 40.3 MGG, ×1000.

Figure 40.4 MGG, ×1000.

Figure 40.5 H&E, ×400.

Figure 40.6 CD20, ×400.

Figure 40.7 CD138, ×400.

Case 41

Figure 41.1 MGG, ×1000.

Figure 41.2 MGG, ×1000.

Figure 41.3 MGG, ×1000.

Figure 41.4 MGG, ×1000.

Figure 41.5 FSC/SSC.

Figure 41.6 CD34/SSC.

Figure 41.7 CD45/SSC.

Case 42

Figure 42.1 H&E, ×100.

Figure 42.2 H&E, ×100.

Figure 42.3 CD24/CD66b.

Figure 42.4 CD24/FLAER.

Figure 42.5 MGG, ×1000.

Figure 42.6 MGG, ×1000.

Figure 42.7 MGG, ×1000.

Figure 42.8 MGG, ×1000.

Figure 42.9 H&E, ×100.

Figure 42.10 H&E, ×500.

Figure 42.11 H&E, ×500.

Case 43

Figure 43.1 MGG, ×1000.

Figure 43.2 MGG, ×1000.

Figure 43.3 MGG, ×1000.

Figure 43.4 MGG, ×1000.

Figure 43.5 H&E, ×400.

Figure 43.6 CD20, ×400.

Case 44

Figure 44.1 MGG, ×500.

Figure 44.2 MGG, ×500.

Figure 44.3 MGG, ×500.

Figure 44.4 MGG, ×500.

Figure 44.5 MGG, ×500.

Figure 44.6 MGG, ×1000.

Figure 44.7 MGG, ×1000.

Figure 44.8 MRI.

Figure 44.9 MRI.

Figure 44.10 H&E, ×100.

Figure 44.11 H&E, ×400.

Figure 44.12 Cam5.2, ×400.

Figure 44.13 Alcian blue, ×400.

Case 45

Figure 45.1 CD45/SSC.

Figure 45.2 CD10/CD20.

Figure 45.3 MGG, ×1000.

Figure 45.4 MGG, ×1000.

Figure 45.5 CD45/SSC.

Figure 45.6 CD10/CD20.

Figure 45.7 CD10/CD34.

Figure 45.8 TdT/CD79a.

Case 46

Figure 46.1 MGG, ×1000.

Figure 46.2 MGG, ×1000.

Figure 46.3 MGG, ×1000.

Figure 46.4 H&E, ×40.

Figure 46.5 H&E, ×400.

Figure 46.6 CD20, ×40.

Figure 46.7 CD20, ×400.

Case 47

Figure 47.1 MGG, ×1000.

Figure 47.2 MGG, ×1000.

Case 48

Figure 48.1 MGG, ×500.

Figure 48.2 MGG, ×1000.

Figure 48.3 MGG, ×1000.

Figure 48.4 FSC/SSC.

Figure 48.5 H&E, ×500.

Figure 48.6 H&E, ×500.

Figure 48.7 CD20, ×500.

Figure 48.8 CD20, ×500.

Figure 48.9 H&E, ×200.

Figure 48.10 CD20, ×200.

Case 49

Figure 49.1 MGG, ×1000.

Figure 49.2 MGG, ×1000.

Figure 49.3 MGG, ×1000.

Figure 49.4 MGG, ×1000.

Figure 49.5 MGG, ×1000.

Figure 49.6 H&E, ×100.

Figure 49.7 H&E, ×500.

Figure 49.8 H&E, ×500.

Figure 49.9 Mast cell tryptase, ×100.

Figure 49.10

CHIC2

FISH studies.

Figure 49.11 MGG, ×500.

Figure 49.12 MGG, ×1000.

Case 50

Figure 50.1 MGG, ×500.

Figure 50.2 MGG, ×1000.

Figure 50.3 MGG, ×1000.

Figure 50.4 MGG, ×1000.

Case 51

Figure 51.1 MGG, ×1000.

Figure 51.2 MGG, ×1000.

Figure 51.3 CD45/SSC.

Figure 51.4 FSC/SSC.

Case 52

Figure 52.1 MGG, ×1000.

Figure 52.2 MGG, ×1000.

Figure 52.3 CD19/SSC.

Figure 52.4 CD138/SSC.

Figure 52.5 FSC/SSC.

Figure 52.6 H&E, ×100.

Figure 52.7 H&E, ×400.

Figure 52.8 CD138, ×400.

Case 53

Figure 53.1 MGG, ×500.

Figure 53.2 MGG, ×500.

Figure 53.3 MGG, ×1000.

Figure 53.4 H&E, ×400.

Figure 53.5 H&E, ×400.

Figure 53.6 CD20, ×200.

Case 54

Figure 54.1 MGG, ×500.

Figure 54.2 MGG, ×1000.

Figure 54.3 MGG, ×1000.

Figure 54.4 MGG, ×1000.

Figure 54.5 MGG, ×1000.

Figure 54.6 H&E, ×40.

Figure 54.7 H&E, ×200.

Figure 54.8 H&E, ×400.

Figure 54.9 CD117, ×200.

Figure 54.10 CD33, ×200.

Figure 54.11 CD11c, ×200.

Figure 54.12 CD15, ×200.

Figure 54.13 Glycophorin C (CD236R), ×200.

Case 56

Figure 56.1 H&E, ×100.

Figure 56.2 H&E, ×400.

Figure 56.3 CD3, ×200.

Figure 56.4 CD4, ×200.

Figure 56.5 CD5, ×200.

Figure 56.6 Ki-67, ×200.

Figure 56.7 MGG, ×500.

Figure 56.8 MGG, ×500.

Figure 56.9 MGG, ×500.

Figure 56.10 MGG, ×500.

Figure 56.11 H&E, ×400.

Figure 56.12 CD3, ×400.

Figure 56.13 FSC/SSC.

Figure 56.14 CD3/HLA-DR.

Figure 56.15 CD2/CD26.

Case 57

Figure 57.1

Figure 57.2

Figure 57.3 H&E, ×40.

Figure 57.4 H&E, ×400.

Figure 57.5 CD123, ×400.

Figure 57.6 CD56, ×400.

Figure 57.7 CD33, ×400.

Figure 57.8 CD4, ×400.

Figure 57.9 MGG, ×500.

Figure 57.10 MGG, ×1000.

Figure 57.11 MGG, ×1000.

Figure 57.12 CD123/SSC.

Figure 57.13 FSC/SSC.

Figure 57.14 CD45/SSC.

Case 58

Figure 58.1 MGG, ×500.

Figure 58.2 MGG, ×1000.

Figure 58.3 MGG, ×1000.

Figure 58.4 CD3/HLA-DR.

Figure 58.5 CD2/CD26.

Figure 58.6 CD7/CD16.

Figure 58.7 CD4/CD8.

Figure 58.8 H&E, ×40.

Figure 58.9 H&E, ×200.

Figure 58.10 CD3, ×100.

Figure 58.11 CD4, ×100.

Case 59

Figure 59.1 MGG, ×500.

Figure 59.2 MGG, ×500.

Figure 59.3 MGG, ×500.

Figure 59.4 MGG, ×500.

Figure 59.5 MGG, ×1000.

Figure 59.6 MGG, ×1000.

Case 60

Figure 60.1 MGG, ×500.

Figure 60.2 MGG, ×1000.

Figure 60.3 CD5/CD20.

Figure 60.4 FMC7/CD23.

Figure 60.5 Kappa/lambda.

Figure 60.6 CD79b/CD22.

Figure 60.7 Cytoplasmic kappa/lambda.

Figure 60.8 CD20/CD5.

Figure 60.9 Kappa/lambda.

Case 61

Figure 61.1 MGG, x1000.

Figure 61.2 MGG, x1000.

Case 62

Figure 62.1 H&E, x100.

Figure 62.2 H&E, x100.

Figure 62.3 H&E, x400.

Figure 62.4 H&E, x400.

Figure 62.5 MGG, x1000.

Figure 62.6 MGG, x1000.

Figure 62.7 MGG, x1000.

Figure 62.8 MGG, x1000.

Figure 62.9 MGG, x1000.

Figure 62.10 MGG, x1000.

Figure 62.11 CD34/SSC.

Figure 62.12 FSC/SSC.

Figure 62.13 H&E, x100.

Figure 62.14 H&E, x500.

Figure 62.15 H&E, x100.

Figure 62.16 H&E, x500.

Case 63

Figure 63.1 MGG, ×1000.

Figure 63.2 MGG, ×1000.

Figure 63.3 Platelet aggregation studies

Case 64

Figure 64.1 MGG, ×500.

Figure 64.2 MGG, ×1000.

Figure 64.3 MGG, ×1000.

Figure 64.4 H&E, ×100.

Figure 64.5 H&E, ×200.

Figure 64.6 CD117, ×100.

Figure 64.7 CD117, ×500.

Figure 64.8 CD117 perivascular, ×500.

Figure 64.9 MGG, ×500.

Figure 64.10 MGG, ×500.

Figure 64.11 tryptase, ×100.

Figure 64.12 tryptase, ×400.

Figure 64.13 CD117/SSC.

Figure 64.14 CD2/CD117 histogram.

Figure 64.15 CD25/CD117 histogram.

Case 65

Figure 65.1 CT.

Figure 65.2 MGG, ×100.

Figure 65.3 MGG, ×500.

Figure 65.4 MGG, ×500.

Figure 65.5 H&E, ×100.

Figure 65.6 Sirius red, ×100.

Figure 65.7 Amyloid P, ×100.

Figure 65.8 Lambda light chains, ×100.

Figure 65.9 H&E, ×400.

Figure 65.10 CD138, ×100.

Case 66

Figure 66.1 MGG, ×1000

Figure 66.2 MGG, ×1000

Figure 66.3 MGG, ×1000

Figure 66.4 MGG, ×1000

Figure 66.5 CD4/CD8

Figure 66.6 CD7/CD16

Figure 66.7 CD4/CD26

Figure 66.8 H&E, ×200

Figure 66.9 H&E, ×400

Figure 66.10 CD4, ×200

Figure 66.11 CD3, ×200

Case 67

Figure 67.1 MGG, ×500.

Figure 67.2 MGG, ×1000.

Figure 67.3 MGG, ×1000.

Figure 67.4 MGG, ×1000.

Figure 67.5 MGG, ×1000.

Figure 67.6 MGG, ×1000.

Figure 67.7 MGG, ×1000.

Figure 67.8 CD45/SSC.

Figure 67.9 Monocyte CD7/CD56.

Case 68

Figure 68.1 MGG, ×500.

Figure 68.2 MGG, ×500.

Figure 68.3 MGG, ×500.

Figure 68.4 MGG, ×500.

Figure 68.5 H&E, ×400.

Figure 68.6 CD56, ×400.

Figure 68.7 Granzyme B, ×400.

Figure 68.8 EBV EBER, ×400.

Figure 68.9 Metaphase cytogenetics.

Figure 68.10 MGG, ×1000.

Figure 68.11 MGG, ×1000.

Figure 68.12 CD56/SSC.

Figure 68.13 FSC/SSC.

Case 69

Figure 69.1 H&E, ×400.

Figure 69.2 CD138, ×400.

Figure 69.3 CD20, ×400.

Figure 69.4 Cyclin D1, ×400.

Figure 69.5 Kappa, ×400.

Figure 69.6 MGG, ×1000.

Figure 69.7 MGG, ×1000.

Figure 69.8 MGG, ×1000.

Figure 69.9 CD45/CD19.

Figure 69.10 CD56/CD38.

Case 70

Figure 70.1 MGG, ×1000.

Figure 70.2 MGG, ×1000.

Figure 70.3 MGG, ×1000.

Case 71

Figure 71.1 MGG, ×500.

Figure 71.2 MGG, ×500.

Figure 71.3 MGG, ×500.

Figure 71.4 MGG, ×500.

Figure 71.5 H&E, ×100.

Figure 71.6 H&E, ×400.

Figure 71.7 CD24/CD66b.

Figure 71.8 CD24/FLAER.

Figure 71.9 Perls stain, ×500.

Case 72

Figure 72.1 MGG, ×1000.

Figure 72.2 MGG, ×1000.

Figure 72.3 MGG, ×10.

Figure 72.4 MGG, ×1000.

Figure 72.5 CT.

Figure 72.6 CT.

Figure 72.7 CT.

Figure 72.8 MGG, ×1000.

Figure 72.9 MGG, ×1000.

Figure 72.10 Karyogram.

Figure 72.11 (a) Pre-treatment, CD34/SSC, CD33/CD7 and CD19/CD2; (b) following treatment course 1, CD34/SSC, CD33/CD7 and CD19/CD2; (c) following treatment course 2, CD34/SSC, CD33/CD7 and CD19/CD2.

Case 73

Figure 73.1 MGG, ×500.

Figure 73.2 MGG, ×500.

Figure 73.3 MGG, ×1000.

Figure 73.4 MGG, ×1000.

Figure 73.5 CD45/SSC.

Figure 73.6 EpCAM/SSC.

Case 74

Figure 74.1 CT.

Figure 74.2 CT (bone windows).

Figure 74.3 CT.

Figure 74.4 MGG, ×1000.

Figure 74.5 MGG, ×1000.

Figure 74.6 MGG, ×1000.

Figure 74.7 CD34/SSC.

Figure 74.8 CD15/CD117.

Figure 74.9 cCD3/MPO.

Figure 74.10 CD2/CD19.

Figure 74.11 H&E, ×400.

Figure 74.12 CD117, ×400.

Figure 74.13 CD15, ×400.

Case 75

Figure 75.1 MGG, ×1000.

Figure 75.2 MGG, ×1000.

Figure 75.3 MGG, ×1000.

Case 76

Figure 76.1 MGG, ×1000.

Figure 76.2 MGG, ×1000.

Figure 76.3 MGG, ×1000.

Figure 76.4 MGG, ×1000.

Figure 76.5 FSC/SSC.

Figure 76.6 CD13/HLA-DR.

Figure 76.7 CD34 histogram.

Figure 76.8 CD14/CD64.

Figure 76.9 CD56/CD38.

Figure 76.10 MGG, ×1000.

Figure 76.11 MGG, ×1000.

Case 77

Figure 77.1 MGG, ×1000.

Figure 77.2 MGG, ×1000.

Figure 77.3 MGG, ×1000.

Figure 77.4 Kappa/lambda.

Figure 77.5 CD20/CD5.

Figure 77.6 CD23/FMC7.

Case 78

Figure 78.1 MGG, ×1000.

Figure 78.2 MGG, ×1000.

Figure 78.3 MGG, ×1000.

Figure 78.4 MGG, ×1000.

Figure 78.5 MGG, ×1000.

Figure 78.6 MGG, ×1000.

Figure 78.7 H&E, ×100.

Figure 78.8 H&E, ×200.

Figure 78.9 H&E, ×300.

Figure 78.10 H&E, ×400.

Figure 78.11 reticulin, ×100.

Figure 78.12 CD34, ×400.

Figure 78.13 CD34, ×100.

Figure 78.14 CD42b, ×400.

Case 79

Figure 79.1 MGG, ×1000.

Figure 79.2 MGG, ×1000.

Figure 79.3 CD19/SSC.

Figure 79.4 kappa/lambda.

Figure 79.5 CD10/HLA-DR.

Figure 79.6 MGG, ×1000.

Figure 79.7 MGG, ×1000.

Case 80

Figure 80.1 MGG, ×1000.

Figure 80.2 MGG, ×1000.

Figure 80.3 MGG, ×1000.

Figure 80.4 MGG, ×1000.

Figure 80.5 CD34/SSC.

Figure 80.6 CD79a/TdT.

Figure 80.7 CD10/CD20.

Figure 80.8 H&E, ×400.

Figure 80.9 H&E, ×400.

Figure 80.10 Reticulin, ×200.

Figure 80.11 CD34, ×400.

Figure 80.12 TdT, ×400.

Figure 80.13 CD10, ×400.

Figure 80.14 CD20, ×400.

Case 81

Figure 81.1 MRI.

Figure 81.2 PET/CT.

Figure 81.3 MGG, ×1000.

Figure 81.4 MGG, ×1000.

Figure 81.5 MGG, ×1000.

Figure 81.6 MGG, ×1000.

Case 82

Figure 82.1 MGG, ×1000.

Figure 82.2 MGG, ×1000.

Figure 82.3 MGG, ×1000.

Figure 82.4 CD45/SSC.

Figure 82.8 cCD3/MPO.

Figure 82.5 CD19/CD2.

Figure 82.6 CD20/CD10.

Figure 82.7 TdT/CD79a.

Figure 82.8 cCD3/MPO.

Figure 82.9 H&E necrosis, ×40.

Figure 82.10 H&E degenerate cells, ×400.

Figure 82.11 H&E viable, ×100.

Figure 82.12 H&E viable, ×400.

Figure 82.13 CD79a, ×400.

Figure 82.14 PAX5, ×400.

Figure 82.15 CD10, ×400.

Figure 82.16 CD20, ×400.

Figure 82.17 TdT, ×400.

Figure 82.18 MPO, ×400.

Case 83

Figure 83.1 MGG, ×500.

Figure 83.2 MGG, ×500.

Figure 83.3 MGG, ×1000.

Figure 83.4 CD3/HLA-DR.

Figure 83.5 CD4/CD8.

Figure 83.6 CD2/CD26.

Figure 83.7 H&E, ×100.

Figure 83.8 H&E, ×400.

Figure 83.9 CD2, ×40.

Figure 83.10 CD3, ×400.

Figure 83.11 CD5, ×400.

Figure 83.12 CD7, ×400.

Case 84

Figure 84.1 MGG, ×1000.

Figure 84.2 MGG, ×1000.

Figure 84.3 MGG, ×1000.

Figure 84.4 MGG, ×1000.

Figure 84.5 CD5/CD20.

Figure 84.6 FMC7/CD23.

Figure 84.7 CD79b/CD22.

Figure 84.8 CD20, ×100.

Figure 84.9 CD23, ×100.

Figure 84.10 H&E, ×100.

Figure 84.11 H&E, ×400.

Figure 84.12 CD20, ×100.

Figure 84.13 CD5, ×100.

Figure 84.14 CD23, ×100.

Figure 84.15 CD3, ×100.

Case 85

Figure 85.1 MGG, ×1000.

Figure 85.2 MGG, ×1000.

Figure 85.3 MGG, ×1000.

Figure 85.4 MGG, ×1000.

Figure 85.5 CT chest at original presentation.

Figure 85.6 CT chest at second presentation.

Figure 85.7 CD34/SSC.

Figure 85.8 CD56/CD38.

Case 86

Figure 86.1 MGG, ×1000.

Figure 86.2 MGG, ×1000.

Figure 86.3 MGG, ×1000.

Case 87

Figure 87.1 CXR.

Figure 87.2 MGG, ×500.

Figure 87.3 MGG, ×1000.

Figure 87.4 MGG, ×1000.

Figure 87.5 MGG, ×1000.

Figure 87.6 MGG, ×1000.

Figure 87.7 cCD3/MPO.

Case 88

Figure 88.1 MGG, ×500.

Figure 88.2 MGG, ×1000.

Figure 88.3 MGG, ×1000.

Figure 88.4 MGG, ×1000.

Case 89

Figure 89.1 MGG, ×1000.

Figure 89.2 MGG, ×1000.

Figure 89.3 MGG, ×1000.

Figure 89.4 MGG, ×1000.

Case 90

Figure 90.1 PET/CT.

Figure 90.2 H&E, ×40.

Figure 90.3 H&E, ×400.

Figure 90.4 H&E, ×400.

Figure 90.5 CD30, ×400.

Figure 90.6 CD15, ×400.

Figure 90.7 EBV (EBER), ×400.

Figure 90.8 PAX5, ×400.

Figure 90.9 CD5, ×400.

Figure 90.10 CD3, ×400.

Case 91

Figure 91.1 MGG, ×1000.

Figure 91.2 MGG, ×1000.

Figure 91.3 MGG, ×1000.

Figure 91.4 MGG, ×1000.

Figure 91.5 CD34/SSC baseline.

Figure 91.6 CD15/CD117 baseline.

Figure 91.7 CD7/CD33 baseline.

Figure 91.8 CD34/SSC post-induction.

Figure 91.9 CD15/CD117 post-induction.

Figure 91.10 CD33/CD7 post-induction.

Case 92

Figure 92.1 CT.

Figure 92.2 MGG, ×1000.

Figure 92.3 MGG, ×1000.

Figure 92.4 H&E, ×400.

Figure 92.5 H&E, ×400.

Figure 92.6 H&E, ×400.

Figure 92.7 H&E, ×400.

Figure 92.8 CD20, ×200.

Figure 92.9 BCL2, ×400.

Figure 92.10 PAX5, ×400.

Figure 92.11 PAX5, ×400.

Case 93

Figure 93.1 Plain X-ray.

Figure 93.2 Plain X-ray.

Figure 93.3 MRI.

Figure 93.4 MRI.

Figure 93.5 H&E, ×40.

Figure 93.6 H&E, ×400.

Figure 93.7 PAS, ×400.

Figure 93.8 CD45, ×400.

Figure 93.9 CD99, ×400.

Figure 93.10 MGG, ×500.

Figure 93.11 MGG, ×500.

Figure 93.12 MGG, ×1000.

Figure 93.13 MGG, ×1000.

Case 94

Figure 94.1 MGG, ×1000.

Figure 94.2 MGG, ×1000.

Figure 94.3 MGG, ×1000.

Figure 94.4 MGG, ×1000.

Figure 94.4 MGG, ×1000.

Figure 94.5 FSC/SSC.

Figure 94.6 CD45/SSC.

Figure 94.7 cCD3/MPO.

Figure 94.8 CD41/CD33.

Figure 94.9 CD61/CD33.

Case 95

Figure 95.1 MGG, ×1000.

Figure 95.2 MGG, ×1000.

Figure 95.3 MGG, ×1000.

Figure 95.4 MGG, ×1000.

Figure 95.5 MGG, ×1000.

Figure 95.6 MGG, ×1000.

Figure 95.7 FSC/SSC basic.

Figure 95.8 CD45/SSC.

Figure 95.9 CD10/CD20 from blast gate.

Figure 95.10 FSC/SSC back gated from CD45 plot.

Case 96

Figure 96.1 MGG, ×1000.

Figure 96.2 MGG, ×1000.

Figure 96.3 H&E, ×40.

Figure 96.4 H&E, ×400.

Figure 96.5 CD20, ×400.

Figure 96.6 CD11c, ×400.

Figure 96.7 CD72, ×400.

Figure 96.8 TRAP, ×400.

Figure 96.9 Annexin 1, ×400.

Figure 96.10 Reticulin, ×400.

Case 97

Figure 97.1 MGG, ×1000.

Figure 97.2 MGG, ×1000.

Figure 97.3 MGG, ×1000.

Figure 97.4 CXR.

Figure 97.5 CT, anteroposterior.

Figure 97.6 CT, lateral.

Figure 97.7 CT, upper chest.

Figure 97.8 CT, abdomen.

Figure 97.9 CD45/SSC.

Figure 97.10 cCD3/TdT.

Figure 97.11 CD5/CD10.

Figure 97.12 CD7/CD16.

Figure 97.13 H&E, ×40.

Figure 97.14 H&E, ×400.

Figure 97.15 CD3, ×200.

Figure 97.16 TdT, ×200

Figure 97.17 CD10, ×200.

Figure 97.18 CD1a, ×200.

Figure 97.19 Reticulin, ×200.

Case 98

Figure 98.1 MGG, ×1000.

Figure 98.2 MGG, ×500.

Figure 98.3 MGG, ×1000.

Figure 98.4 H&E, ×500.

Figure 98.5 H&E, ×200.

Figure 98.6 H&E, ×400.

Figure 98.7 CD79a, ×200.

Figure 98.8 CD20, ×200.

Figure 98.9 CD10, ×200.

Figure 98.10 BCL2, ×200.

Figure 98.11 MUM1, ×200.

Figure 98.12 Ki-67, ×200.

Case 99

Figure 99.1 CT.

Figure 99.2 CT.

Figure 99.3 CT.

Figure 99.4 MGG, ×1000.

Figure 99.5 MGG, ×1000.

Figure 99.6 MGG, ×1000.

Figure 99.7 MGG, ×1000.

Figure 99.8 CD5/CD20.

Figure 99.9 FMC7/CD23.

Figure 99.10 Kappa/lambda.

Figure 99.11 H&E, ×400.

Figure 99.12 H&E, ×400.

Figure 99.13 CD20, ×400.

Figure 99.14 CD10, ×400.

Figure 99.15 BCL2, ×400.

Figure 99.16 Ki-67, ×200.

Case 100

Figure 100.1 MGG, ×1000.

Figure 100.2 MGG, ×1000.

Figure 100.3 MGG, ×1000.

Figure 100.4 MGG, ×1000.

Figure 100.5 CD138/SSC.

Figure 100.6 FSC/SSC.

Figure 100.7 CD19/cytoplasmic kappa.

Figure 100.8 CD19/cytoplasmic lambda.

Figure 100.9 MGG, ×500.

Figure 100.10 MGG, ×500.

Figure 100.11 MGG, ×1000.

Figure 100.12 MGG, ×1000.

Figure 100.13 H&E, ×400.

Figure 100.14 CD138, ×400.

Figure 100.15 CD20, ×200.

Figure 100.16 CD30, ×400.

Figure 100.17 MUM1, ×400.

Figure 100.18 ALK, ×400.

Figure 100.19 FISH for ALK at 2p23.

Practical Flow Cytometry in Haematology: 100 Worked Examples

Mike Leach FRCP, FRCPath

Consultant Haematologist and Honorary Senior LecturerHaematology Laboratories and West of Scotland Cancer CentreGartnavel General HospitalGlasgow, UK

Mark Drummond PhD, FRCPath

Consultant Haematologist and Honorary Senior LecturerHaematology Laboratories and West of Scotland Cancer CentreGartnavel General HospitalGlasgow, UK

Allyson Doig MSc, FIBMS

Haemato-Oncology Laboratory ManagerHaematology LaboratoriesGartnavel General HospitalGlasgow, UK

Pam McKay

Consultant HaematologistHaematology Laboratories and West of Scotland Cancer CentreGartnavel General HospitalGlasgow, UK

Bob Jackson

Consultant PathologistDepartment of PathologySouthern General HospitalGlasgow, UK

Barbara J. Bain MBBS, FRACP, FRCPath

Professor of Diagnostic HaematologySt Mary's Hospital Campus of Imperial CollegeFaculty of Medicine, Londonand Honorary Consultant Haematologist,St Mary's Hospital, London, UK

This edition first published 2015 © 2015 by John Wiley & Sons, Ltd

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Library of Congress Cataloging-in-Publication Data

Leach, Richard M. (Haematologist), author.

Practical flow cytometry in haematology : 100 worked examples / Mike Leach [and 5 others].

p. ; cm.

Includes index.

ISBN 978-1-118-74703-2 (hardback)

I. Title.

[DNLM: 1. Hematologic Diseases–diagnosis–Case Reports. 2. Hematologic Neoplasms–diagnosis–Case Reports. 3. Flow Cytometry–methods–Case Reports. 4. Hematology–methods–Case Reports. WH 120]

RC636

616.1′5075—dc23

2015007734

A catalogue record for this book is available from the British Library.

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Preface

In our first publication ‘Practical Flow Cytometry in Haematology Diagnosis’ we presented an outline approach to the use and applications of flow cytometric immunophenotyping in the diagnostic haematology laboratory. We showed how this technique could be used to study blood, bone marrow and tissue fluid samples in a variety of clinical scenarios to achieve a diagnosis, taking into account important features from the clinical history and examination alongside haematology, morphology, biochemistry, immunology, cytogenetic, histopathology and molecular data. This text was illustrated with a series of ‘worked examples’ from real clinical cases presenting to our institution. These cases have proven to be very popular and so a companion publication dedicated to 100 new ‘worked examples’ seemed justified and is presented here.

The principles used in the approach to each case are exactly the same as used in the first publication and cases are illustrated with tissue pathology and cytogenetic and molecular data, which are integrated to generate, where appropriate, a diagnosis based on the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. We present a spectrum of clinical cases encountered in our department from both adult and paediatric patients and of course, if the title is to be justified, flow cytometry plays a role in every case. Furthermore, we present both neoplastic and reactive disorders and the cases appear in no particular order so that the reader should have no pre-conceived idea as to the nature of the diagnosis in any case. May−Grünwald−Giemsa (MGG)-stained films of peripheral blood and bone marrow aspirates are presented with flow cytometry data alongside haematoxylin and eosin (H&E)-stained bone marrow and tissue biopsy sections. Immunohistochemistry is used to further clarify the tissue lineage and cell differentiation. Cytogenetic studies using metaphase preparations are used to identify translocations and chromosome gains and losses whilst interphase fluorescence in situ hybridisation (FISH) studies and polymerase chain reaction (PCR) are used to identify gene fusions, break-aparts and deletions. The presentation is brought to a conclusion and the particular features that are important in making a diagnosis are highlighted and discussed. The cases are also listed according to disease classification toward the end (page 390) so that the text can also be used as a reference manual.