Flow Cytometry: First Principles

2nd ed, by Alice Longobardi Givan, 273 pp, with illus, New York, NY, Wiley-Liss, 2001.This concise and well-organized text provides a good introduction tothe basic principles of flow cytometry, assuming no prior knowledge of the field. The first several chapters are devoted to a description of flow cytometric systems, including instrumentation, sample preparation, and data analysis. This is followed by chapters describing applications of the system in clinical laboratories, as well as research areas. Chapters are supplemented with ample references for more in-depth exposure to topics, and there is a thorough glossary of terms provided at the end of the book.The first chapter gives a brief history of the development of flow cytometry, followed in the next chapter by an explanation of the differences between high-tech research instruments (multilaser with sorting capabilities) and more clinically oriented bench-top instruments. The emphasis in the reminder of the text is on flow principles that apply to both types of instruments.The third chapter explains the basics of instrumentation, including lasers, fluidics, signal detection, and electronics. This explanation is followed by a chapter on computerized data storage and analysis, and a chapter describing lasers in more detail, as well as fluorochromes, mirrors, and filters.The remaining 7 chapters of the book are devoted to the various applications of flow cytometric systems. The sixth chapter is concerned with leukocytes, characterization of their surface antigens by flow, and the principles of cell gating. The next chapter focuses on characterization of intracellular proteins, with emphasis on technical pitfalls. The eighth and ninth chapters provide good introductions to the topics of DNA analysis and flow cytometric cell sorting.An overview of flow cytometric applications in clinical medicine is given in the 10th chapter. This overview includes descriptions of leukemia/lymphoma immunophenotyping, human immunodeficiency virus monitoring, reticulocyte counting, platelet analysis, stem cell enumeration, and fetal hemoglobin analysis. DNA analysis in surgical pathology and use of the flow cross-match in solid organ transplantation are also covered.The 11th chapter turns to research applications of flow cytometry. The reader is given a sampling of the diversity of uses for this technology in fields like cellular immunology, cell kinetics, aquatic biology, microbiology, molecular biology, reproductive biology, and biochemistry. Finally, the brief 12th chapter offers some speculations on the future of flow cytometry.In general, the writing is clear and concise, with effective use of figures and tables to convey complex concepts. The fifth chapter on lasers, fluorochromes, and filters is particularly well done. On the other hand, several complex subjects receive such concise coverage that they may be difficult for the beginner to comprehend. For example, Figure 3.9 and the accompanying text on electronics need clarification. Similarly, Figures 4.3 and 4.4 (histograms, dot plots, and contour plots) and Figure 8.8 (effect of cell clumps on DNA analysis) are not very clearly explained. Dot-plot axes in Figure 7.2 are mislabeled.Compared to the 1st edition of this text, the current edition provides more information on fluorochrome and laser options for multicolor analysis, a discussion of apoptosis, a new chapter on intracellular protein staining, and an expanded section on sorting. In summary, this book can provide students, technologists, and clinicians with a good basic introduction to flow cytometry and a guide to the subject literature.