Flow Cytometry : First Principles

Flow cytometry continually amazes scientists with its ever-expanding utility. Advances in flow cytometry have opened new directions in theoretical science, clinical diagnosis, and medical practice. The new edition of __Flow Cytometry: First Principles__ provides a thorough update of this now classic text, reflecting innovations in the field while outlining the fundamental elements of instrumentation, sample preparation, and data analysis. __Flow Cytometry: First Principles__, Second Edition explains the basic principles of flow cytometry, surveying its primary scientific and clinical applications and highlighting state-of-the-art techniques at the frontiers of research. This edition contains extensive revisions of all chapters, including new discussions on fluorochrome and laser options for multicolor analysis, an additionalsection on apoptosis in the chapter on DNA, and new chapters onintracellular protein staining and cell sorting, including high-speed sorting and alternative sorting methods, as well as traditional technology. This essential resource: * Assumes no prior knowledge of flow cytometry * Progresses with an informal, engaging lecture style from simpleto more complex concepts * Offers a clear introduction to new vocabulary, principles of instrumentation, and strategies for data analysis * Emphasizes the theory relevant to all flow cytometry, with examples from a variety of clinical and scientific fields __Flow Cytometry: First Principles__, Second Edition provides scientists, clinicians, technologists, and students with the knowledge necessary for beginning the practice of flow cytometry and for understanding related literature.Content: Chapter 1 The Past as Prologue (pages 1–9): Chapter 2 Setting the Scene (pages 11–14): Chapter 3 Instrumentation: Into the Black Box (pages 15–39): Chapter 4 Information: Harnessing the Data (pages 41–57): Chapter 5 Seeing the Light: Lasers, Fluorochromes, and Filters (pages 59–80): Chapter 6 Cells from Without: Leukocytes, Surface Proteins, and the Strategy of Gating (pages 81–113): Chapter 7 Cells from Within: Intracellular Proteins (pages 115–122): Chapter 8 Cells from Within: DNA in Life and Death (pages 123–158): Chapter 9 The Sorting of Cells (pages 159–174): Chapter 10 Disease and Diagnosis: The Clinical Laboratory (pages 175–194): Chapter 11 Out of the Mainstream: Research Frontiers (pages 195–223): Chapter 12 Flowing On: The Future (pages 225–228): Contents......Page 6 Preface......Page 9 Acknowledgments (First Edition)......Page 12 Acknowledgments (Second Edition)......Page 14 Index......Page 16 1 The Past as Prologue......Page 27 2 Setting the Scene......Page 36 3 Instrumentation:Into the Black Box......Page 40 ILLUMINATION OF THE STREAM......Page 41 CENTERING CELLS IN THE ILLUMINATING BEAM......Page 43 DETECTION OF SIGNALS FROM CELLS......Page 50 ELECTRONICS......Page 56 DATA STORAGE......Page 65 DATA ANALYSIS......Page 67 GENERAL THEORY......Page 82 LASERS......Page 84 FLUOROCHROMES......Page 88 PARTITIONING THE SIGNAL WITH LENSES, FILTERS, AND MIRRORS......Page 95 SPECTRAL COMPENSATION......Page 99 FURTHER READING......Page 103 6 Cells from Without: Leukocytes, Surface Proteins, and the Strategy of Gating......Page 104 CELLS FROM BLOOD......Page 105 STAINING FOR SURFACE MARKERS......Page 110 CONTROLS......Page 113 QUANTITATION......Page 118 SENSITIVITY......Page 121 THE STRATEGY OF GATING......Page 122 GATING ON FLUORESCENCE......Page 131 7 Cells from Within: Intracellular Proteins......Page 137 METHODS FOR PERMEABILIZING CELLS......Page 138 EXAMPLES OF INTRACELLULAR STAINING......Page 140 FLUOROCHROMES FOR DNA ANALYSIS......Page 145 PLOIDY......Page 147 CELL CYCLE ANALYSIS......Page 153 TWO-COLOR ANALYSIS FOR DNA AND ANOTHER PARAMETER......Page 164 CHROMOSOMES......Page 169 APOPTOSIS......Page 172 NECROSIS......Page 176 FURTHER READING......Page 179 SORTING THEORY......Page 181 CHARACTERIZATION OF SORTED CELLS......Page 188 ALTERNATIVE METHODS FOR SORTING......Page 192 THE CONDITION OF CELLS AFTER SORTING......Page 194 10 Disease and Diagnosis: The Clinical Laboratory......Page 197 THE HEMATOLOGY LABORATORY......Page 200 THE PATHOLOGY LABORATORY......Page 208 SOLID ORGAN TRANSPLANTATION......Page 211 COMMENTS......Page 214 11 Out of the Mainstream: Research Frontiers......Page 217 FUNCTIONAL ASSAYS......Page 218 THE AQUATIC ENVIRONMENT......Page 224 REPORTER MOLECULES......Page 229 MICROBIOLOGY......Page 233 MOLECULAR BIOLOGY......Page 235 MULTIPLEX CYTOMETRY FOR SOLUBLE ANALYTES......Page 240 REPRODUCTIVE TECHNOLOGY......Page 242 12 Flowing On: The Future......Page 246 BOOKS......Page 250 CATALOGUES......Page 252 MISCELLANEOUS......Page 253 Glossary......Page 255 Figure Credits......Page 277 Contents 6 Preface 9 Acknowledgments (First Edition) 12 Acknowledgments (Second Edition) 14 Index 16 1 The Past as Prologue 27 2 Setting the Scene 36 3 Instrumentation:Into the Black Box 40 ILLUMINATION OF THE STREAM 41 CENTERING CELLS IN THE ILLUMINATING BEAM 43 DETECTION OF SIGNALS FROM CELLS 50 ELECTRONICS 56 4 Information: Harnessing the Data 65 DATA STORAGE 65 DATA ANALYSIS 67 5 Seeing the Light: Lasers, Fluorochromes, and Filters 82 GENERAL THEORY 82 LASERS 84 FLUOROCHROMES 88 PARTITIONING THE SIGNAL WITH LENSES, FILTERS, AND MIRRORS 95 SPECTRAL COMPENSATION 99 FURTHER READING 103 6 Cells from Without: Leukocytes, Surface Proteins, and the Strategy of Gating 104 CELLS FROM BLOOD 105 STAINING FOR SURFACE MARKERS 110 CONTROLS 113 QUANTITATION 118 SENSITIVITY 121 THE STRATEGY OF GATING 122 GATING ON FLUORESCENCE 131 7 Cells from Within: Intracellular Proteins 137 METHODS FOR PERMEABILIZING CELLS 138 EXAMPLES OF INTRACELLULAR STAINING 140 8 Cells from Within: DNA in Life and Death 145 FLUOROCHROMES FOR DNA ANALYSIS 145 PLOIDY 147 CELL CYCLE ANALYSIS 153 TWO-COLOR ANALYSIS FOR DNA AND ANOTHER PARAMETER 164 CHROMOSOMES 169 APOPTOSIS 172 NECROSIS 176 FURTHER READING 179 9 The Sorting of Cells 181 SORTING THEORY 181 CHARACTERIZATION OF SORTED CELLS 188 ALTERNATIVE METHODS FOR SORTING 192 THE CONDITION OF CELLS AFTER SORTING 194 10 Disease and Diagnosis: The Clinical Laboratory 197 THE HEMATOLOGY LABORATORY 200 THE PATHOLOGY LABORATORY 208 SOLID ORGAN TRANSPLANTATION 211 COMMENTS 214 11 Out of the Mainstream: Research Frontiers 217 FUNCTIONAL ASSAYS 218 THE AQUATIC ENVIRONMENT 224 REPORTER MOLECULES 229 MICROBIOLOGY 233 MOLECULAR BIOLOGY 235 MULTIPLEX CYTOMETRY FOR SOLUBLE ANALYTES 240 REPRODUCTIVE TECHNOLOGY 242 12 Flowing On: The Future 246 General References 250 BOOKS 250 CATALOGUES 252 MISCELLANEOUS 253 Glossary 255 Figure Credits 277 Flow cytometry continually amazes scientists with its ever-expanding utility. Advances in flow cytometry have opened new directions in theoretical science, clinical diagnosis, and medical practice. The new edition of Flow Cytometry: First Principles provides a thorough update of this now classic text, reflecting innovations in the field while outlining the fundamental elements of instrumentation, sample preparation, and data analysis.Flow Cytometry: First Principles, Second Edition explains the basic principles of flow cytometry, surveying its primary scientific and clinical applications and highlighting state-of-the-art techniques at the frontiers of research. This edition contains extensive revisions of all chapters, including new discussions on fluorochrome and laser options for multicolor analysis, an additional section on apoptosis in the chapter on DNA, and new chapters on intracellular protein staining and cell sorting, including high-speed sorting and alternative sorting methods, as well as traditional technology. The Past As Prologue -- Setting The Scene -- Instrumentation : Into The Black Box -- Information : Harnessing The Data -- Seeing The Light : Lasers, Fluorochromes, And Filters -- Cells From Without : Leukocytes, Surface Proteins, And The Strategy Of Gating -- Cells From Within : Intracellular Proteins -- Cells From Within : Dna In Life And Death -- The Sorting Of Cells -- Disease And Diagnosis : The Clinical Laboratory -- Out Of The Mainstream : Research Frontiers -- Flowing On : The Future. Alice Longobardi Givan. Previous Ed.: 1992. Includes Bibliographical References And Index. Also Available In An Electronic Version.