CONTENTS Preface iv PART ONE: The Principles of Hematology 1 1 Safety and Quality in the Hematology Laboratory................................. 1 An Overview of the Hematology Laboratory 1 The Study of Hematology 1 Functions of the Hematology Laboratory 1 Safety in the Hematology Laboratory 2 The Safety Officer 2 Occupational Safety and Health Administration Acts and Standards 2 Avoiding Transmission of Infectious Diseases 3 Immune Status: Screening and Vaccination 5 Safe Work Practices and Protective Techniques for Infection Control 6 Safety Manual, Policies, and Practices 6 Standard Precautions 6 Handwashing 6 Personal Protective Equipment 7 Decontamination of Work Surfaces, Equipment, and Spills 8 General Infection Control Safety Practices 9 OSHA Medical Waste Standards 11 Quality Assessment in the Hematology Laboratory 12 Regulations and Organizations Impacting Quality 12 Components of Quality Assessment 12 Nonanalytical Factors in Quality Assessment 12 Quality Control in the Hematology Laboratory 16 Terms Used in Clinical Quality Control 16 Functions of a Quality Control Program 17 Using Statistical Analysis of Results in Quality Assessment 18 Other Statistical Applications in the Hematology Laboratory 20 CHAPTER HIGHLIGHTS 20 REVIEW QUESTIONS 21 BIBLIOGRAPHY 23 2 Principles of Blood Collection................ 25 Quality in Phlebotomy 25 Quality Assessment 25 Patient Care Partnership 25 The Phlebotomist as Laboratory Ambassador 26 Patients with Special Considerations 26 Pediatric Patients 26 Adolescent Patients 26 Geriatric Patients 26 Blood Collection Supplies and Equipment 26 Anticoagulants 26 Adverse Effects of Additives 27 Safe Blood Collection 27 Evacuated Blood Collection Tubes 29 Anticoagulants and Additives in Evacuated Blood Tubes 31 Capillary Blood 31 Specimen Handling Requirements 32 CHAPTER HIGHLIGHTS 45 REVIEW QUESTIONS 47 REFERENCE 48 BIBLIOGRAPHY 48 3 Molecular Genetics and Cellular Morphology....50 Cellular Ultrastructure and Organization 50 Cellular Membranes 50 Cell Volume Homeostasis 52 Reactive and Neoplastic Growth Processes 53 Cytoplasmic Organelles and Metabolites 53 Cellular Inclusions and Metabolites 54 Nuclear Characteristics 54 Chromosomes 56 Activities of the Nucleus 58 The Foundations of Genetic Interactions 60 Genetic Alterations 61 Molecular Techniques in Hematology 63 Minimal Residual Disease 63 Hematopathology 67 Gene Rearrangement Studies 69 CHAPTER HIGHLIGHTS 69 REVIEW QUESTIONS 70 BIBLIOGRAPHY 72 4 Hematopoiesis............................. 73 Hematopoiesis Defined 73 Origin of Blood Cells 73 Types of Human Stem Cells 73 Early Development of Blood Cells 73 Bone Marrow Sites and Function 75 Cellular Elements of Bone Marrow 76 Progenitor Blood Cells 76 Erythropoiesis 79 Granulopoiesis 79 Lymphopoiesis 79 Megakaryopoiesis 79 Other Cells Found in Bone Marrow 79 Interleukins 79 Hematopoietic Growth Factors 82 Examination of Maturing Blood Cells 83 General Cellular Characteristics 83 Nuclear Characteristics 83 Cytoplasmic Characteristics 85 Mature Blood Cells in Peripheral Blood 86 CHAPTER HIGHLIGHTS 86 REVIEW QUESTIONS 87 BIBLIOGRAPHY 87 PART TWO: Erythrocytes 89 5 Erythrocyte Maturation, Physiology, and Lifecycle.............................. 89 Erythropoiesis 90 Erythropoietin 90 General Characteristics of Maturation and Development 91 Developmental Stages 92 Reticulocytes 94 vi
Contents vii Disorders Related to Erythrocyte Maturation and Production 97 Disorders of Erythropoietin 97 Red Cell Increases 98 Defective Nuclear Maturation 98 Characteristics and Biosynthesis of Hemoglobin 98 Genetic Inheritance of Hemoglobin 98 Chemical Composition and Configuration of Hemoglobin 98 The Role of 2,3-Diphosphoglycerate 99 Oxygen Dissociation and Alterations 99 Carbon Dioxide Transport 100 Biosynthesis of Hemoglobin 101 Disorders Related to Hemoglobin Biosynthesis 104 Disorders of Heme (Porphyrin) Synthesis 104 Disorders of Iron Metabolism 105 Disorders of Globulin Synthesis 108 Ontogeny of Hemoglobin 108 Variant Forms of Normal Hemoglobin 109 Abnormal Hemoglobin Molecules 110 Analysis of Hemoglobin 110 Membrane Characteristics and Metabolic Activities of Erythrocytes 111 Membrane Characteristics 112 Cytoplasmic Characteristics 113 Metabolic Activities 113 Catabolism of Erythrocytes 114 Measurement of Erythrocytes 116 Mean Corpuscular Volume 116 Mean Corpuscular Hemoglobin 117 Mean Corpuscular Hemoglobin Concentration 117 CHAPTER HIGHLIGHTS 118 CASE STUDIES 119 REVIEW QUESTIONS 121 BIBLIOGRAPHY 124 6 Erythrocyte Morphology and Inclusions...... 126 Erythrocytes: Normal and Abnormal 126 Types of Variations in Erythrocyte Size 126 Kinds of Variations in Erythrocyte Shape 127 Alterations in Erythrocyte Color 134 Varieties of Erythrocyte Inclusions 135 Alterations in Erythrocyte Distribution 137 Parasitic Inclusions in Erythrocytes 137 Malaria 137 Other Parasitic Inclusions 141 CHAPTER HIGHLIGHTS 142 CASE STUDY 142 REVIEW QUESTIONS 143 BIBLIOGRAPHY 144 7 Classification and Laboratory Assessment of Anemias................................. 145 Causes of Anemia 145 Clinical Signs and Symptoms of Anemia 145 Classification of Anemias 145 Laboratory Assessment of Anemias 146 Quantitative Measurements of Anemia 147 Semiquantitative Grading of Erythrocyte Morphology 147 Supplementary Assessment of Anemias 148 CHAPTER HIGHLIGHTS 148 REVIEW QUESTIONS 148 BIBLIOGRAPHY 149 8 Acute and Chronic Blood Loss Anemias....... 150 Acute Blood Loss Anemia 150 Etiology 150 Physiology 150 Laboratory Findings 150 Chronic Blood Loss Anemia 150 Etiology 150 CHAPTER HIGHLIGHTS 151 CASE STUDIES 151 REVIEW QUESTIONS 153 BIBLIOGRAPHY 153 9 Aplastic and Related Anemias............... 154 Aplastic Anemia 154 Etiology 154 Pathophysiology 155 Clinical Features 158 Laboratory Findings 158 Treatment 158 Congenital Red Blood Cell Related Disorders 159 Telomeres 159 Laboratory Findings in Bone Marrow Failure Syndromes 160 Pure Red Cell Aplasia 160 Diamond-Blackfan Anemia 160 Fanconi Anemia 161 Transient Erythroblastopenia of Childhood 162 Congenital Dyserythropoietic Anemia 163 CHAPTER HIGHLIGHTS 163 CASE STUDIES 163 REVIEW QUESTIONS 165 BIBLIOGRAPHY 165 10 Iron Deficiency Anemia and Anemia of Chronic Inflammation............................ 166 Scope of the Problem 166 Iron Deficiency Anemia 166 Early Diagnosis 166 Etiology 166 Epidemiology 167 Physiology 168 Pathophysiology 169 Clinical Signs and Symptoms 170 Laboratory Characteristics 170 Anemia of Inflammation or Anemia of chronic Disorders 173 Etiology 173 Pathophysiology 173 Laboratory Characteristics 174 Treatment 175 CHAPTER HIGHLIGHTS 175 CASE STUDIES 176 REVIEW QUESTIONS 179 BIBLIOGRAPHY 180 11 Megaloblastic Anemias..................... 181 Megaloblastic Anemias 181 Etiology 181 Epidemiology 182 Physiology 182 Vitamin B 12 (Cobalamin) Transport 182 Vitamin B 12 (Cobalamin) and Folic Acid Deficiencies 183 Pathophysiology 184 Gastric Pathological Findings 184 Clinical Signs and Symptoms 186 Laboratory Findings 186 Treatment and Monitoring Therapy 188 CHAPTER HIGHLIGHTS 188 CASE STUDY 190 REVIEW QUESTIONS 191 BIBLIOGRAPHY 191
viii Contents 12 Hemolytic Anemias........................ 192 Hemolytic Anemias 192 Inherited Hemolytic Anemia 192 Acquired Hemolytic Anemia 197 Pathophysiology 200 Diagnostic Tests 201 Paroxysmal Nocturnal Hemoglobinuria 201 Etiology 201 Epidemiology 202 Pathophysiology 202 Clinical Signs and Symptoms 202 Laboratory Findings 203 Treatment 203 Paroxysmal Cold Hemoglobinuria 203 CHAPTER HIGHLIGHTS 203 CASE STUDIES 204 REVIEW QUESTIONS 207 BIBLIOGRAPHY 208 13 Hemoglobinopathies and Thalassemias....... 210 Hemoglobin Defects 210 Demographics 210 Etiology 210 Sickle Cell Disease 211 Etiology 212 Epidemiology 212 Pathophysiology 212 Clinical Signs and Symptoms 213 General Signs and Symptoms 214 Laboratory Testing 214 Special Laboratory Testing 215 Management of Sickle Cell Disease 216 Sickle Cell Syndromes: Pathogenesis and New Approaches 218 Sickle b-thalassemia 218 Sickle-C Disease 219 Sickle Cell Trait 219 Thalassemia 220 Demographics 220 Etiology 220 Pathophysiology 220 b-thalassemia 220 α-thalassemia 223 Other Hemoglobinopathies 224 Hemoglobin C Disease 224 Hemoglobin SC Disease 224 Hemoglobin D Disease 224 Hemoglobin E Disease 224 Hemoglobin H Disease 225 Methemoglobinemia 225 Unstable Hemoglobins 225 Hereditary Persistance of Fetal Hemoglobin 225 CHAPTER HIGHLIGHTS 226 CASE STUDIES 227 REVIEW QUESTIONS 232 BIBLIOGRAPHY 233 PART THREE: Leukocytes 235 14 Leukocytes: The Granulocytic and Monocytic Series................................... 235 Introduction 235 The Granulocytic Series 235 Production of Neutrophils, Eosinophils, and Basophils 235 Sites of Development and Maturation 236 Development and Proliferation of Neutrophils, Eosinophils, and Basophils 236 Distribution of Neutrophils, Eosinophils, and Basophils 236 Normal Maturational Characteristics of Neutrophils, Eosinophils, and Basophils 237 Mature Forms 239 Granulation in Mature Forms 239 The Monocytic-Macrophage Series 240 Production and Development of Monocytes and Macrophages 240 Morphological Characteristics 241 Reference Ranges of Granulocytes and Monocytes 241 Functional Properties of Granulocytes and Monocytes 242 General Characteristics 243 The Role of Macrophages 243 Acute Inflammatory Response 243 Sepsis 245 Steps in Phagocytosis 245 Specialized Functions of Granulocytes 247 Assessment Methods 248 Total Leukocyte Count 248 Differential Blood Smear Evaluation 248 Absolute Cell Counts 248 Erythrocyte Sedimentation Rate 248 Assessment of Eosinophils and Basophils 249 Leukocyte Alkaline Phosphatase Test 249 Neutrophilic Function 249 Neutrophilic Hypersegmentation Index 249 CHAPTER HIGHLIGHTS 249 CASE STUDY 250 REVIEW QUESTIONS 251 BIBLIOGRAPHY 253 15 Nonmalignant Disorders of Granulocytes and Monocytes............................... 254 Quantitative Disorders 254 Leukocytosis 254 Leukocytopenia 255 Morphological Abnormalities of Mature Granulocytes 256 Toxic Granulation 256 Döhle Bodies 256 Hypersegmentation 256 Pelger-Huët Anomaly 256 May-Hegglin Anomaly 257 Chédiak-Higashi Syndrome 257 Alder-Reilly Inclusions 258 Ehrlichia 258 Abnormalities of Mature Granulocytes in Body Fluids 258 Qualitative Disorders 258 Defective Locomotion and Chemotaxis 258 Defects in Microbicidal Activity 259 Other Functional Anomalies of Neutrophils 259 Monocyte-Macrophage Disorders 259 Gaucher Disease 259 Niemann-Pick Disease 260 CHAPTER HIGHLIGHTS 260 CASE STUDIES 260 REVIEW QUESTIONS 264 BIBLIOGRAPHY 265 16 Leukocytes: Lymphocytes and Plasma Cells.... 266 Anatomical Origin and Development of Lymphocytes 266 Sites of Lymphocytic Development 266 Lymphocyte Physiology 267 Normal Reference Values 267 Determining Absolute Lymphocyte Values 268 Morphological Characteristics of Normal Lymphocytes 268 Maturational Stages 269 Specific Lymphocyte Morphological Variations 272
Contents ix Characteristics of Lymphocytes 273 Major Lymphocyte Categories and Functions 273 Monoclonal Antibodies 274 Major Lymphocyte Membrane Characteristics and Development 274 Functional Testing of Lymphocytes 277 Plasma Cell Development and Maturation 277 Plasma Cell Development 277 Maturational Morphology 277 Plasmacytoid Lymphocytes 277 Plasma Cell Disorders 278 CHAPTER HIGHLIGHTS 278 REVIEW QUESTIONS 279 BIBLIOGRAPHY 280 17 Leukocytes: Nonmalignant Lymphocytic Disorders................................ 281 Characteristics of Lymphocytes 281 Lymphocytosis 281 Disorders Associated with Lymphocytosis 282 Infectious Mononucleosis 282 Cytomegalovirus Infection 286 Toxoplasmosis 287 Infectious Lymphocytosis 288 Bordetella pertussis (Haemophilus pertussis) Infection 289 Lymphocytopenia 289 Immune Disorders Associated with Lymphocytopenia 289 DiGeorge Syndrome 289 Acquired Immunodeficiency Syndrome(HIV/AIDS) 289 Systemic Lupus Erythematosus 293 CHAPTER HIGHLIGHTS 294 CASE STUDIES 294 REVIEW QUESTIONS 297 BIBLIOGRAPHY 297 18 Characteristics of Leukemias, Lymphomas, and Myelomas................................ 298 Comparison of Leukemias, Lymphomas, and Myelomas 298 Forms of Leukemia 298 Classifications of Leukemias 299 French-American-British Classification 299 World Health Organization Classification 299 Prognosis and Treatment 300 Factors Related to the Occurrence of Leukemia 300 Genetic and Immunological Factors 300 Occupational Exposure 302 Environmental Exposure 302 Chemical and Drug Exposure 302 Genetic Abnormalities and Associations 302 Viral Agents 303 Secondary Causes of Leukemias 303 Demographic Distribution of Leukemia and Lymphomas 304 Ethnic Origin and Race 304 Gender 305 CHAPTER HIGHLIGHTS 305 REVIEW QUESTIONS 306 BIBLIOGRAPHY 306 19 Acute Leukemias.......................... 307 Introduction 307 Epidemiology of Acute Leukemias 307 Prognosis of Acute Leukemias 308 French-American-British and World Health Organization Categories 308 French-American-British (FAB) Classification 308 World Health Organization (WHO) Organization Classfication 308 General Characteristics of Acute Myeloid Leukemias 309 Genetic Differences 309 Micro-RNAs 310 Acute Myeloid Leukemia 310 FAB M0 311 Acute Myeloid Leukemia (FAB M1) 311 Acute Myeloid Leukemia (FAB M2) 312 Acute Promyelocytic Leukemia (FAB M3) 312 Acute Myelomonocytic Leukemia (FAB M4) 313 Acute Monocytic Leukemia (FAB M5) 314 Erythroleukemia (FAB M6) 315 Selected Examples of Unusual Forms 316 Acute megakaryoblastic leukemia FAB M7 316 Epidemiology of Acute Lymphoblastic Leukemia 316 Prognosis of Acute Lymphoblastic Leukemia 317 Classifications of Acute Lymphoblastic Leukemia 317 Characteristics of Acute Lymphoblastic Leukemia 317 Clinical Signs and Symptoms 317 Laboratory Data 317 Special Identification Techniques 318 Treatment 319 Mixed Lineage Leukemia 319 Cytogenetic Analysis 319 Cytogenetic Analysis in Acute Myeloid Leukemia 319 Cytogenetics in Acute Lymphoblastic Leukemia 320 Principles of Special Cytochemical Stains 320 Sudan Black B Stain 321 Myeloperoxidase Stain 326 Periodic Acid-Schiff Stain 322 Esterase Stains 323 Phosphatase Stains 324 Monoclonal Antibodies 325 Immunophenotyping 325 Lymphoid 325 Other Surface Membrane Markers 326 Life-Threatening Emergencies 326 Infection 326 Bleeding 326 Leukemic Infiltration of Organs 326 Metabolic Abnormalities 326 Hyperleukocytosis 327 Treatment Options 327 Relapse 328 Stem Cell Transplant 328 Future Trends Vaccines 328 CHAPTER HIGHLIGHTS 328 CASE STUDIES 329 REVIEW QUESTIONS 333 BIBLIOGRAPHY 335 20 Lymphoid and Plasma Cell Neoplasms........ 337 Mature B-Cell Neoplasms Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma 337 Epidemiology 337 Etiology 337 Cytogenetics 338 Molecular Genetics 339 Staging and Prognosis 339 Clinical Signs and Symptoms 340 Laboratory Data 341 Treatment Options 341 Minimal Residual Disease 342 Hairy Cell Leukemia 342 Hairy Cell Leukemia Variant 343 Prolymphocytic Leukemia 344
x Contents Multiple Myeloma (Plasma Cell Myeloma) 344 Epidemiology 344 Etiology 344 Clinical Signs and Symptoms 344 Laboratory Data 344 Treatment 345 Waldenström Primary Macroglobulinemia (Lymphoplasmacytic lymphoma) 346 Epidemiology 346 Etiology 346 Clinical Signs and Symptoms 346 Laboratory Data 346 Treatment 347 Lymphomas 347 Relationship Between Lymphomas and Leukemias 347 Characteristics 347 Categories 347 Pathophysiology 349 Precursors of Hodgkin Disease and B-Cell Lymphomas 350 Hodgkin Disease 350 Non-Hodgkin Lymphoma 351 Characteristics of Other Forms 352 Sézary Syndrome 353 Mature T-Cell and NK-Cell Neoplasms 353 CHAPTER HIGHLIGHTS 353 CASE STUDIES 354 REVIEW QUESTIONS 358 BIBLIOGRAPHY 358 PART FOUR: Myeloproliferative Neoplasms 361 21 Myeloproliferative Neoplasms............... 361 General Characteristics of Myeloproliferative Neoplasms 362 Relationship of the Myeloproliferative Neoplasms 362 Common Disorders of Hemostasis and Coagulation 362 Prognosis and Treatment 363 Chronic Myelogenous Leukemia 363 Epidemiology 363 Pathophysiology 363 Cytogenetics 364 Genetic Alterations 364 Laboratory Data 365 Cytogenetic Studies 367 Prognosis and Treatment 367 Minimal Residual Disease 368 Leukemia-Specific Targets 369 Allogeneic Bone Marrow Transplantation 370 Polycythemia Vera, Essential Thrombocytosis (Essential Thrombocythemia), and Primary Myelofibrois 370 Polycythemia Vera 370 Epidemiology 370 Etiology 370 Pathophysiology 370 Karyotype 371 Clinical Signs and Symptoms 371 Laboratory Data 371 Abnormalities of Hemostasis and Coagulation 372 Other Laboratory Assays 372 Treatment 373 Prognosis and Complications 374 Primary Myelofibrosis 374 Epidemiology 374 Pathophysiology 374 Karyotype 375 Clinical Signs and Symptoms 375 Cellular Alterations 375 Prognosis 376 Treatment 376 Essential Thrombocytosis/Essential Thrombocythemia 376 Diagnostic Characteristics 376 Epidemiology 377 Pathophysiology 377 Karyotype 377 Clinical Signs and Symptoms 377 Laboratory Findings 377 Relationship of Thrombocythemia and PV 377 Treatment 378 CHAPTER HIGHLIGHTS 378 CASE STUDIES 379 REVIEW QUESTIONS 382 BIBLIOGRAPHY 384 22 Myelodysplastic Syndromes and Myelodysplastic/ Myeloproliferative Neoplasms............... 386 Classification 386 Myelodysplastic Syndromes 386 Myelodysplastic/myeloproliferative neoplasms 386 Pathophysiology 387 Myelodysplastic Syndrome 387 Etiology 388 Epidemiology 389 Chromosomal Abnormalities 389 Clinical Signs and Symptoms 390 Laboratory Manifestations 390 Features of Selected Types of Myelodysplastic Syndromes 391 Myelodysplastic/Myeloproliferative Neoplasms 392 Chronic myelomonocytic leukemia 392 Other Classifications 393 Treatment Strategies 393 Relationship of Karyotype to Prognosis 394 CHAPTER HIGHLIGHTS 395 CASE STUDIES 396 REVIEW QUESTIONS 397 BIBLIOGRAPHY 398 PART FIVE: Principles and Disorders of Hemostasis and Thrombosis 399 23 Principles of Hemostasis and Thrombosis..... 399 Overview of Hemostasis and Thrombosis 400 Blood Vasculature: Structure and function 400 Arteries and Veins 400 Arterioles and Venules 400 Capillaries 400 Vasculature Physiology 401 The Role of Vasoconstriction in Hemostasis 401 The Role of the Endothelium 401 Maintenance of Vascular Integrity 402 The Megakaryocytic Cell Series 403 General Characteristics of Megakaryocytic Development 403 Cellular Ultrastructure of a Mature Platelet 405 Platelet Kinetics, Life Span, and Normal Values 406 Platelet Function in Hemostasis 406 Overall Functions of Platelets 406 Platelet Adhesion 406 Platelet Aggregation 407 Platelet Plug Consolidation and Stabilization 408 Laboratory Assessment of Platelets 408
Contents xi Blood Coagulation Factors 410 Basic Concepts of Blood Coagulation 410 Common Characteristics of Coagulation Factors 410 Characteristics of Individual Factors 410 The Mechanism of Coagulation 413 Coagulation Pathways 413 Thrombin-Mediated Reactions 414 Fibrinolysis 414 Other Systems and Inhibitors 415 Laboratory Assessment of Blood Coagulation Factors 416 International Normalized Ratio 416 Specialized Assays for Coagulation Factors 417 Anticoagulants 417 New Thromboplastins 418 Assays for Fibrin Formation 419 Normal Protective Mechanisms Against Thrombosis 420 Normal Blood Flow 420 Removal of Activated Clotting Factors and Particulate Material 420 The Natural Anticoagulant Systems 420 Cellular Regulators 424 Modern View of Hemostasis 424 CHAPTER HIGHLIGHTS 424 REVIEW QUESTIONS 426 BIBLIOGRAPHY 429 24 Disorders of Hemostasis and Thrombosis..... 431 Vascular Disorders 431 Abnormal Platelet Morphology 432 Quantitative Platelet Disorders 432 Thrombocytopenia 432 Thrombocytosis 436 Qualitative Characteristics of Platelets: Thrombocytopathy 436 Types of Platelet Dysfunctions 437 Bleeding Disorders Related to Blood Clotting 439 Defective Production 440 Disorders of Destruction and Consumption 445 Disorders Related to Elevated Fibrin Split Products 449 The Hypercoagulable State 450 Primary States of Hypercoagulability 450 Secondary States of Hypercoagulability 450 Pregnancy-Associated Thrombosis 450 General Features 451 Impaired Fibrinolysis 454 Antithrombin III Deficiency 456 Laboratory Assessment of Hypercoagulable States 457 CHAPTER HIGHLIGHTS 458 CASE STUDIES 460 REVIEW QUESTIONS 462 BIBLIOGRAPHY 463 PART SIX: Fundamentals of Hematological Analysis 465 25 Body Fluid Analysis........................ 465 Introduction 466 Cerebrospinal Fluid 466 Anatomy and Physiology 466 Production of Cerebrospinal Fluid 466 Specimen Collection: Lumbar Puncture 466 Laboratory Analysis 467 Pleural, Peritoneal, and Pericardial Fluids 473 Effusions: Transudates and Exudates 473 Pleural Fluid 473 Peritoneal Fluid 478 Pericardial Fluid 482 Seminal Fluid 484 Synovial Fluid 486 Anatomy and Physiology of Joints 486 Purpose of Arthrocentesis 486 Aspiration 487 Laboratory Assays 487 Body Fluid Slide Preparation 490 Staining of Body Fluid Sediment 490 Amniotic Fluid 490 CHAPTER HIGHLIGHTS 490 REVIEW QUESTIONS 493 BIBLIOGRAPHY 496 26 Manual Procedures in Hematology........... 498 Procedural Format 498 Coagulation Procedures 516 Specimen Quality 516 Special Collection Techniques 516 Anticoagulants 517 Specimen Handling 517 Specimen Preparation 517 General Sources of Error 517 Quality Control 518 REVIEW QUESTIONS 522 27 Instrumentation in Hematology............. 526 Instrumental Principles 527 The Electrical Impedance Principle 527 The Optical Detection Principle 527 Characteristics of Light Scatter 527 Radio Frequency 528 Fundamentals of Laser Technology 528 Principles of Flow Cytometry 529 The Basis of Cellular Identification 529 Whole Blood Cell Analysis 531 Types of Automated Cell Counting Instruments 532 General Histogram Characteristics 543 Analysis of Instrumental Data Output 543 The Erythrocyte Histogram 543 Quantitative Descriptors of Erythrocytes 543 The Leukocyte Histogram 544 Platelet Histograms 545 Derived Platelet Parameters 546 Laser Technology 547 Red Blood Cells/Platelets 547 Peroxidase 548 Basophil/Lobularity (Nuclear) Channel 549 Lymphocyte Subtyping 549 Applications of Flow Cytometry 549 General Properties of Flow Cytometry 550 Hematological Applications 550 Clinical Applications of Flow Cytometry 550 Other Cellular Applications 551 Digital Microscopy 554 Artificial Neural Networks 554 Digital Cell Morphology 554 Instruments in Coagulation Studies 555 Electromechanical Methods 555 Photo-Optical Methods 555 Viscosity-based Detection System 558 Platelet Agglutination 559 Platelet Aggregation 559 New Automation 559 Summary 559 Instrumental Principles 559 Analysis of Electrical Impedance Instrumental Data Output 559
xii Contents Laser Technology 560 Applications of Flow Cytometry 560 Instruments in Coagulation Studies 560 CASE STUDIES 560 REVIEW QUESTIONS 569 BIBLIOGRAPHY 571 APPENDICES A Answers to Review Questions............... 573 B The Basics of Medical Terminology.......... 578 C SI Units................................. 580 D English-Spanish Medical Phrases for the Phlebotomist............................. 581 E MSDS Sheet.............................. 582 F Tube Guide.............................. 588 G Frequently Used Abbreviations.............. 590 Glossary 591 Index 602