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Methods in cell biology, vol. 33: Flow cytometry
JOURNAL OF EXPERIMENTAL MARINE BIOLOGY AND ECOLOGY
ELSEVIER
J. Exp. Mar. Biol. Ecol. 175 (1993) 133-136
Book Reviews Methods iti Cell Biology, Vol. 33: Flow Cytometry, edited by Zbigniew Darzynkiewicz & Harry A. Crissman; Harcourt Brace Jovanovich, Academic Press, London; 1990; 716 pp.; GBP 33.00 (comb bound), GBP 63.00 (case bound); ISBN O-12-203050-8 (comb bound), O-12-564133-8 (case bound). The growing popularity of flow cytometry has recently stimulated the publication of a rash of beginner’s guides to the subject: thankfully, this volume is not one of them. It weighs in at over 700 densely printed comb-bound pages, and consists of 54 chapters authored by 87 contributors. The list includes many who pioneered the development of flow cytometry or opened up new applications in cell biology, and they waste no space on drawings of flow cells or descriptions of lasers. Instead, much of the book is devoted to short chapters which introduce specific measuring techniques and present carefully documented protocols, complete with reagent lists and recommended standards, to enable readers to carry them out in their own laboratory. Topics covered range from well established techniques for determining nucleic acid content and cell cycle progression to tentative methods for a range of other parameters such as membrane potential, intracellular pH, calcium and enzymes. There is a summary of the spectral properties of most fluorochromes used in flow cytometry, and several articles on the analysis of isolated nuclei and chromosomes. Rcadcrs of this journal are likely to be particularly interested in the use of flow cytometry in the environmental sciences, and to seek guidance on issues which currently hinder wider adoption of the technique. Answers to one set of questions, concerning the high cost of instruments and their reputation for being difficult to adjust and deploy (especially on board ship) can only be provided by instrument designers and manufacturers and are outwith the scope of this book. Another area of concern is the degree to which staining and data processing techniques developed using a rather restrictcd set of mammalian cells can be transferred to other cell types. Hcrc, the book is probably best considered as an invaluable short-cut to the literature and as a source of inspiration for workers who are devising their own cytometric methods. In this respect, there are five chapters near the end of the volume which are devoted to nonmammalian cells and are particularly welcome. One is a short article by Steen, Skarstad & Boye on DNA determinations in bacteria: this is mainly concerned with Eschevichitr coli cultures, whose cells are much larger than the bacteria which are likely to be cncountered in aquatic ecosystems. Nevertheless, this chapter provides useful pointers to appropriate staining techniques and instrument design. The technically uninitiated may be interested to discover that the installation of multiwatt lasers as illumination sources does not necessarily produce high sensitivity instruments. Techniques for examining Elscwcr SSDl
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plant protoplasts (Galbraith) and isolated plant nuclei (Bergounioux & Brown) provide useful starting points for those working on phytoplankton cells, even though differences in morphology and cell structure may prevent them from being directly applicable. Finally, a substantial chapter by Clarice Yentsch follows a different format from the rest of the volume, making a clear case for the exciting potential of flow cytometry in oceanographic research and providing a masterly summary of the state of the art in 1989, when the book went to press. The breadth of the material covered does make the presentation somewhat breathless, however, and this chapter is full of one-line references to new developments that really deserve a page of explanation. There are also areas of methodology which are either missing or inadequately discussed, including the use of simplified excitation spectra in indicating algal groups, the potential of multi-angle light scattering for identifying specific cell types and cluster morphologies, and the great power of combining optical signals with Coulter-style electronic volume measurements. The book as a whole pays much more attention to optimising preparative chemistry and fluorochrome staining than to the design of optical configurations and statistical methods of data analysis. Overall, Elow Cytometry represents a major editorial achievement and a significant contribution to the subject. Environmental scientists will feel the need for a similarly comprehensive set of protocols for analysing such interesting and varied cell types as aquatic bacteria, microheterotrophs, coccolithophorids and cyanobacteria. This volume may inadvertently provide them with a stimulus by showing how much work has still to be done. Alex Cunningham Department of Physics and Applied University of Strathclyde John Anderson Building 107 Rottenrow Glasgow, G4 ONG UK
Physics
Plankton Regulution Dynamics - Experiments and Models in Rotifer Corltinuous Cultures,
Walz; Springer-Verlag, Heidelberg, New York; 1993; 308 pp.; (EcoW. D. Billings, G. Galley, 0. L. Lange, J. S. Olsen & DM 248; ISBN 3-540-55955-g (Heidelberg), O-387-55955-8 (New York).
edited by Norbert
logical Studies, Vol. 98. Eds.:
H. Remmert);
The continuous culture of micro-organisms, in this case rotifers, which is the subject of this book is certainly ready for an overview, not only to bring together the considerable information gained from the various applications of this approach, but to enlighten the newcomer to this fascinating technique for dealing with dynamic metazoan predator-prey systems. The main contributors to this book have all had a great deal of experience with rotifer continuous cultures and deal competently with the subject, ranging from the theoretical considerations to the application of the technique to aquaculture. The
ELSEVIER
J. Exp. Mar. Biol. Ecol. 175 (1993) 133-136
Book Reviews Methods iti Cell Biology, Vol. 33: Flow Cytometry, edited by Zbigniew Darzynkiewicz & Harry A. Crissman; Harcourt Brace Jovanovich, Academic Press, London; 1990; 716 pp.; GBP 33.00 (comb bound), GBP 63.00 (case bound); ISBN O-12-203050-8 (comb bound), O-12-564133-8 (case bound). The growing popularity of flow cytometry has recently stimulated the publication of a rash of beginner’s guides to the subject: thankfully, this volume is not one of them. It weighs in at over 700 densely printed comb-bound pages, and consists of 54 chapters authored by 87 contributors. The list includes many who pioneered the development of flow cytometry or opened up new applications in cell biology, and they waste no space on drawings of flow cells or descriptions of lasers. Instead, much of the book is devoted to short chapters which introduce specific measuring techniques and present carefully documented protocols, complete with reagent lists and recommended standards, to enable readers to carry them out in their own laboratory. Topics covered range from well established techniques for determining nucleic acid content and cell cycle progression to tentative methods for a range of other parameters such as membrane potential, intracellular pH, calcium and enzymes. There is a summary of the spectral properties of most fluorochromes used in flow cytometry, and several articles on the analysis of isolated nuclei and chromosomes. Rcadcrs of this journal are likely to be particularly interested in the use of flow cytometry in the environmental sciences, and to seek guidance on issues which currently hinder wider adoption of the technique. Answers to one set of questions, concerning the high cost of instruments and their reputation for being difficult to adjust and deploy (especially on board ship) can only be provided by instrument designers and manufacturers and are outwith the scope of this book. Another area of concern is the degree to which staining and data processing techniques developed using a rather restrictcd set of mammalian cells can be transferred to other cell types. Hcrc, the book is probably best considered as an invaluable short-cut to the literature and as a source of inspiration for workers who are devising their own cytometric methods. In this respect, there are five chapters near the end of the volume which are devoted to nonmammalian cells and are particularly welcome. One is a short article by Steen, Skarstad & Boye on DNA determinations in bacteria: this is mainly concerned with Eschevichitr coli cultures, whose cells are much larger than the bacteria which are likely to be cncountered in aquatic ecosystems. Nevertheless, this chapter provides useful pointers to appropriate staining techniques and instrument design. The technically uninitiated may be interested to discover that the installation of multiwatt lasers as illumination sources does not necessarily produce high sensitivity instruments. Techniques for examining Elscwcr SSDl
Scicncc 0 0 2 2
B.V.. Amsterdam 0 9 8
I(9
3 ) E 0
14
8 - R
134
Book Reviebt:r / J. Exp. Mur. Bid. Ed.
I75 (1994) 133-1.36
plant protoplasts (Galbraith) and isolated plant nuclei (Bergounioux & Brown) provide useful starting points for those working on phytoplankton cells, even though differences in morphology and cell structure may prevent them from being directly applicable. Finally, a substantial chapter by Clarice Yentsch follows a different format from the rest of the volume, making a clear case for the exciting potential of flow cytometry in oceanographic research and providing a masterly summary of the state of the art in 1989, when the book went to press. The breadth of the material covered does make the presentation somewhat breathless, however, and this chapter is full of one-line references to new developments that really deserve a page of explanation. There are also areas of methodology which are either missing or inadequately discussed, including the use of simplified excitation spectra in indicating algal groups, the potential of multi-angle light scattering for identifying specific cell types and cluster morphologies, and the great power of combining optical signals with Coulter-style electronic volume measurements. The book as a whole pays much more attention to optimising preparative chemistry and fluorochrome staining than to the design of optical configurations and statistical methods of data analysis. Overall, Elow Cytometry represents a major editorial achievement and a significant contribution to the subject. Environmental scientists will feel the need for a similarly comprehensive set of protocols for analysing such interesting and varied cell types as aquatic bacteria, microheterotrophs, coccolithophorids and cyanobacteria. This volume may inadvertently provide them with a stimulus by showing how much work has still to be done. Alex Cunningham Department of Physics and Applied University of Strathclyde John Anderson Building 107 Rottenrow Glasgow, G4 ONG UK
Physics
Plankton Regulution Dynamics - Experiments and Models in Rotifer Corltinuous Cultures,
Walz; Springer-Verlag, Heidelberg, New York; 1993; 308 pp.; (EcoW. D. Billings, G. Galley, 0. L. Lange, J. S. Olsen & DM 248; ISBN 3-540-55955-g (Heidelberg), O-387-55955-8 (New York).
edited by Norbert
logical Studies, Vol. 98. Eds.:
H. Remmert);
The continuous culture of micro-organisms, in this case rotifers, which is the subject of this book is certainly ready for an overview, not only to bring together the considerable information gained from the various applications of this approach, but to enlighten the newcomer to this fascinating technique for dealing with dynamic metazoan predator-prey systems. The main contributors to this book have all had a great deal of experience with rotifer continuous cultures and deal competently with the subject, ranging from the theoretical considerations to the application of the technique to aquaculture. The