Flow Cytometry: First principles

Flow Cytometry: First principles

Journal of Immunological Methods, 165 (1993) 135 135 © 1993 Elsevier Science Publishers B.V. All rights reserved 0022-1759/93/$06.00 JIM06835 Book...

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Journal of Immunological Methods, 165 (1993) 135


© 1993 Elsevier Science Publishers B.V. All rights reserved 0022-1759/93/$06.00


Book review

Longobardi Givan, A., Flow Cytometry: First Principles, xiv + 202 pp., illus. Wiley-Liss, New York, 1992. Soft cover (price not listed) ISBN 0-471-56095-2 As explained in the preface, this book is based on a course given by the author at Newcastle University in England for new and potential flow cytometer users with various professional backgrounds including researchers, technologists, and clinicians. The diversity of readers at whom it is aimed reflects the multiple applications of this technique as well as the different skills involved in using it. The text is divided into ten chapters, with the subject matter evolving from the general to the particular. Chapters 1 and 2 review the historical development of flow cytometry, from the origins of fluorescence microscopy and Kamentsky's spectrophotometric cell sorter to modern, commercially produced flow cytometers. Chapter 3 describes the main instrument components - the optical bench, the flow system, electronics, and cell sorting controls; their construction and operation are depicted with the aid of diagrams. In Chapter 4 different ways of storing the recorded data are shown and methods for their analysis discussed. Chapter 5 deals in detail with the optical aspects of flow cytometry. After explaining the theoretical basis of fluorescence it describes the generation of an illuminating beam, the emission of light signals by the fluorochromestained cells and their registration by photodetectors, and the correction for any spectral overlap between fluorochromes. Chapter 6 focuses on the cytometric analysis of lymphocytes. Procedures are outlined for staining these cells with fluorochrome-coupled monoclonal antibodies specific for surface antigens characteristic of defined lymphocyte subpopulations. A strategy is elucidated for gating, i.e., selecting cells according to their fluorescence/scatter properties and thus restrict-

ing the data in further analyses. Chapter 7 contains protocols for analyzing cellular DNA. It lists suitable fluorochromes and indicates approaches to the quantitation of diploid DNA and cell cycle analysis, described techniques for analyzing both DNA and another parameter such as RNA or protein markers, then characterizing chromosomes, and finally monitoring cell death by staining with propidium iodide. Chapter 8 is devoted to the uses of flow cytometry in clinical diagnosis in the haematology, immunology, oncology, and pathology laboratories. Particular emphasis is placed on the classification of leukaemias and lymphomas according to their cell-membrane markers, and its correlation with the clinical course as followed by DNA analysis and BrdU incorporation. Another noteworthy example is the use as an aid in selecting transplantation patients of the flow cytometric crossmatch assay for recipient's immunoglobulin bound to the surface of the donor's T cells. Chapter 9 shows some of the more unusual applications of flow technology, e.g., in measuring changes in the intracellular calcium level, or the gel microdroplet method for studying bacterial cell division. In the last chapter the author projects her thoughts on the future prospects of flow cytometry which include an acquired capability to analyze particles as small as viruses, mitochondria, etc. and to determine simultaneously several parameters. The text is followed by general references to previously published books on flow cytometry, catalogues of instrument parts and chemicals used, and other sources of information on the subject. A helpful glossary of technical expressions and a subject index are appended. The readability and compactness of this book contribute to its suitability as laboratory manual for biologists and biomedical scientists engaged in cell analysis a n d / o r sorting. F. BOREK