Diagnostic Flow Cytometry in Cytology

✍ Scribed by Pranab Dey

• Publisher: Springer
• Year: 2021
• Tongue: English
• Leaves: 201
• Category: Library

✦ Synopsis

The book covers the essential practical techniques of flow cytometry in detail. It is divided into two sections: The first section includes the basic practical techniques of flow cytometry in cytology samples. Chapters under this section provide detailed description of the sampling technique, processing, acquisition of the sample, instrumentation and basic principles of flow cytometry. The second section elucidates clinical applications of flow cytometry. Chapters cover the flow cytometry applications in various haematolymphoid neoplasms, tumors of solid organs and body fluid samples. The flow cytometry findings of different tumors are described with the help of multiple colored cytology microphotographs, flow cytometry graphs, boxes, and tables. In addition, it also describes other ancillary techniques in those neoplastic lesions.

The book helps practicing pathologists, technical staff and post graduate students to understand flow cytometry findings of the haematolymphoid neoplasms and solid tumor with special emphasis on cytology along with advanced technique. This book will help the students to interpret flow cytometry graphs.

✦ Table of Contents

Preface
Acknowledgements
Contents
About the Author
Abbreviations
Part I: Practical Aspects of Flow Cytometry
1: Introduction and History of Flow Cytometry
1.1 Introduction
1.2 History of Flow Cytometry
1.2.1 Early Motivation
1.2.2 Counting the Flowing Cells
1.2.3 Haematology Sample
1.2.4 Differential Count in the Blood Sample
1.2.5 Flow Sorter
1.3 Fluorescence Stain in the Flow Cytometer
1.4 Further Improvements in Flow Cytometry
References
2: Basic Principles and Instrumentation of Flow Cytometry
2.1 Introduction
2.2 Principles of Flow Cytometry
2.2.1 The Fluidics System
2.3 Optical System
2.4 Fluorescence Emission
2.4.1 Collection of Light
2.4.2 Optical Filters [2]
2.4.3 Electronics System
2.4.4 Computer System
2.4.5 Flow Cytometric Cell Sorting
References
3: Sample Preparation and Data Acquisition in Flow Cytometry
3.1 Introduction
3.2 Basic Requirements
3.3 Cytology Samples for Flow Cytometry
3.4 Sample Collection
3.5 Single-Cell Preparation
3.5.1 Limitations of the Enzymatic Method
3.6 Fixation
3.7 Permeabilization
3.8 RBC Lysing Solution
3.9 Staining
3.9.1 DNA Flow Cytometry [2]
3.9.2 Control
3.10 Data Acquisitions
References
4: Display and Interpretation of Data in Flow Cytometry
4.1 Distribution of Fluorescence Intensity
4.2 Gating
4.2.1 The Crucial Gating in Flow Cytometry
4.3 Backgating
References
5: Quality Control in Flow Cytometry
5.1 Introduction
5.2 Internal Quality Control
5.3 Instrument Quality Control
5.4 The Critical Factors to Have Good Quality FCM Data
5.4.1 Sensitivity
5.5 PMT Voltage Setting
5.5.1 Compensation
5.6 Daily Cytometer Set up
5.7 External Quality Assessment (EQA)
References
6: Fluorescent Probes and Different Useful Markers for Flow Cytometry
6.1 Staining by the Fluorochrome Dye
6.1.1 Applications of FRET
6.1.2 The Desirable Characteristic of a Fluorochrome Dye
6.1.3 Fluorochrome Dyes Used in a Flow Cytometer
6.1.3.1 Single Fluorochrome Dye
6.1.3.2 Tandem Dyes
Precautions of Using Tandem Dye
6.1.3.3 Quantum Dots(QD)
Advantages of QD
6.1.3.4 Reporter Molecules
6.1.4 Multicoloured Flow Cytometry
6.1.5 Basic Principles of Panel Design
6.1.5.1 Machine Understanding
References
7: Nuclei Acid Dye and DNA Content Measurement in Flow Cytometry
7.1 Types of DNA Dye
7.2 Description of Different DNA Dyes
7.2.1 Intercalator Dyes
7.2.2 Minor Groove Binding Dye
7.2.3 Bis-Intercalator Dyes
7.3 DNA Content and Ploidy Analysis
7.4 Standard Nomenclature
7.4.1 Control Diploid Population
7.4.2 Staining for DNA FCM [4]
7.4.2.1 Materials
7.5 Data Acquisition
7.6 Interpretation
References
Part II: Diagnostic Applications of Flow Cytometry in Cytology
8: Classification of Lymphoma, Different Markers and Approach
8.1 Non-Hodgkin Lymphoma
8.2 Markers of Lymphoid Cell Lineage
8.2.1 NK Cell
8.3 Hodgkin Lymphomas (HL)
8.4 Limitations
8.5 Approach to Flow Cytometry of Lymph Node
8.6 Cytology Smear and Panel of Antibody
References
9: Markers for Immunophenotyping in Flow Cytometry
9.1 Introduction
9.2 CD Markers
9.2.1 CD2
9.2.1.1 Diagnostic Applications
9.2.2 CD3
9.2.2.1 Diagnostic Applications
9.2.3 CD4
9.2.3.1 Diagnostic Applications
9.2.4 CD8
9.2.4.1 Diagnostic Applications
9.2.5 CD5
9.2.5.1 Diagnostic Applications
9.2.6 CD7
9.2.6.1 Diagnostic Applications
9.2.7 CD10
9.2.7.1 Diagnostic Applications
9.2.8 CD11B
9.2.8.1 Diagnostic Applications
9.2.9 CD14
9.2.9.1 Diagnostic Applications
9.2.10 CD15
9.2.10.1 Diagnostic Applications
9.2.11 CD19
9.2.11.1 Diagnostic Applications
9.2.12 CD20
9.2.12.1 Diagnostic Applications
9.2.13 CD23
9.2.13.1 Diagnostic Applications
9.2.14 CD25
9.2.14.1 Diagnostic Applications
9.2.15 CD30
9.2.15.1 Diagnostic Applications
9.2.16 CD38
9.2.16.1 Diagnostic Applications
9.2.17 CD43
9.2.17.1 Diagnostic Applications
9.2.18 CD45
9.2.18.1 Diagnostic Applications
9.2.19 CD56
9.2.19.1 Diagnostic Applications
9.2.20 CD79a
9.2.20.1 Diagnostic Applications
9.2.21 CD103
9.2.22 CD117
9.2.22.1 Diagnostic Applications
9.2.23 CD138
9.2.23.1 Diagnostic Applications
9.3 Other Markers Used in Lymphoma
9.3.1 Terminal Deoxynucleotidyl Transferase (TdT)
9.3.1.1 Diagnostic Applications
9.3.2 HLA-DR
9.3.2.1 Diagnostic Applications
9.3.3 PAX5
9.3.3.1 Diagnostic Applications
References
10: Detection of Lymphoma: Clonality Demonstration by Flow Cytometry
10.1 Introduction
10.2 Clonal Proliferation of B Cells
10.2.1 Light Chain Restriction
10.2.2 Aberrant Expression of Certain Antigen
10.3 Immature B Cells
10.3.1 Reactive Lymph Node
10.3.2 B Cell Lymphoma with no Light Chain Expression
10.3.3 Clonal Proliferation of T Cells
10.3.4 Aberrant Expression or Loss of T Cell Antigen
10.3.5 Abnormality of CD4 and CD8 Expression
10.3.6 Increased Forward Scatter
10.3.7 The Expression of Other Markers
10.3.8 Presence of Markers of Blasts
References
11: Flow Cytometry of B-Non Hodgkin Lymphoma
11.1 Introduction
11.2 Diagnosis of Individual NHL
11.2.1 Small Lymphocytic Lymphoma (SLL)
11.2.2 Mantle Cell Lymphoma (MCL)
11.2.3 Follicular Lymphoma (FL)
11.2.4 Marginal Zone Lymphoma (MZL)
11.2.5 Lymphoplasmacytic Lymphoma (LPL)
11.3 Lymphomas of Large-Sized Cells
11.3.1 Diffuse Large B-Cell Lymphoma (DLBCL)
11.3.2 Burkitt Lymphoma (BL)
11.3.3 Hairy Cell Leukaemia (HCL)
11.4 Immature B Cell
11.4.1 B-Lymphoblastic Lymphoma
11.5 Plasma Cell Neoplasm
11.5.1 Differential Diagnosis
11.6 CD5 Positive B-Cell Lymphomas
11.6.1 CD10 Positive Lymphomas
11.7 CD5 and CD10 Negative Lymphoma
References
12: Flow Cytometry of Mature and Immature T-Cell Lymphoma
12.1 Introduction
12.2 Mature T-Cell Lymphomas
12.3 Mycosis Fungoides and Sezary Syndrome
12.4 Peripheral T-Cell Lymphoma (PTCL)
12.5 Angio-Immunoblastic T-Cell Lymphoma (AITL)
12.6 Hepatosplenic T-Cell Lymphoma (HSTL) [5]
12.7 Extranodal Natural Killer/T-Cell Lymphoma
12.8 Immature T-Cell Lymphoma
12.8.1 T-Cell Lymphoblastic Leukaemia (T-LBL) or Lymphoma (T-ALL) [6]
References
13: Flow Cytometry of Body Cavity Fluid
13.1 Introduction
13.2 Detection of Malignancy in Fluid
13.3 DNA Flow Cytometry
13.3.1 Immunophenotyping to Detect Malignancy
13.3.2 Precautions to Take for the Best Result
13.3.3 Possible Pitfalls
13.4 Detection of Lymphoma in Fluid
13.4.1 Panel of Markers in Leukaemia/Lymphoma
13.4.2 Diagnostic Features
13.5 Primary Effusion Lymphoma (PEL)
13.6 Urine Flow Cytometry
References
14: Flow Cytometry of Solid Tumours
14.1 Introduction
14.2 Advantages of Flow Cytometry in the Detection of Carcinoma
14.3 Diagnosis of the Small Round Cell Tumours
14.4 DNA Content Analysis and Synthetic Phase Assessment
14.5 Limitation of DNA Analysis by FCM
14.6 The Response of Cancer Chemotherapeutic Drugs
14.7 Expression of Oncogene Markers and Receptor Expression
References
15: Self-Assessment Test in Flow Cytometry
15.1 Answer Key of Chap. 15