- Email: [email protected]
Shedding light on living cells
TIBS 15-MARCH1990 such as analysis of gene expression and structure, or the isolation of mitochondriai DNA, but the emphasis is on the particular difficulties posed by plants - the abundance of nucleases for instance, or the need to break the cell wall. Other chapters cover topics unique to plants and include chloroplast molecular biology, transformation with Agrobacterium and protoplast preparation. The book ends with chapters on three specialized subjects: plant viruses, Chlamydomonas and cyanobacteria, all of which, although amenable to most techniques, nonetheless have their own particular vagaries, about which it is useful to know. Throughout the book, the usual format of the 'Practical Approach' series is taken, with tables of detailed step-by-step protocols interspersed in the more discursive text. The latter, although not essential in order to follow the procedures, provides the rationale behind the techniques, and also considers the limitations, so that adapting them for your own system is much easier. One
aspect that is different from most of the earlier books in the series is that it is available in spiralbound format - much more 'practical' for the bench than paperback. The intended audience for Methods in Plant Molecular Biology is quite different. It is specifically designed for use as a manual for an undergraduate laboratory course, and is based on one which is run at the University of Illinois. The experiments described introduce a wide range of different techniques, starting from basic cloning and analysis of DNA, through translation in vitro by isolated chloroplasts and the wheat germ system, and concluding with dideoxynucleotide DNA sequencing and transformation with Agrobacterium. This course is meant to be done two days a week over a 15-week semester, but it is possible to pick out small groups of experiments to illustrate more specific techniques, such as basic cloning or characterization of a genomic DNA clone, which could be done in shorter periods. Each experiment has an introduction which explains the overall
procedure, and the rationale for doing it. The protocols themselves are clearly presented and easy to follow, and include interesting snippets of information, such as why chloroform is often included in a phenol extraction, or why different proteins give different responses with the Lowry test. In addition, there are highlighted warnings to alert the students to potential dangers to themselves, or to their experimental results, and notes for instructors to facilitate the setting up and running of the practicals. Its value, however, is likely to be confined to someone thinking of instituting a practical course in this area, rather than as a research laboratory manual. In the end, whichever book you buy, I bet that within a short period of time you will be developing your own way of doing things - and then you can write your own 'definitive' manual!
Shedding light on living cells
the light microscope, is being used increasingly to study the dynamics of cellular processes. This renaissance of the light microscope has come about partly because of the development of techniques that facilitate the non-invasive observation of live cells. Phase and Nomarski optics convert small refractive index differences within a sample into intensity changes. More recently, electronic contrast enhancement has pushed the limits of what can be detected by Nomarski optics to the extent that individual microtubules with a diameter of one-twentieth the wavelength of visible light may be clearly discerned in favourable circumstances. Powerful though these techniques are, they are undiscriminating in that all structures that have a different refractive index from their immediate environment will be imaged. Stains with specific affinities have long been used as a method of discriminating a structure of interest from a complex background in fixed cells. Fluorescent stains have become increasingly used because of the high detection sensitivities that may be obtained. Immunofluorescence techniques have been developed to the extent that a specific stain may be constructed for practically any cellular component that possesses a unique hapten. In recent years a number of research groups have developed methods for using selective fluorescent staining techniques in living cells. These methods open up the exciting possibility of being able to follow
the fate of a particular cellular component during normal cellular processes. Furthermore, fluorochromes have been developed that alter their fluorescence emission characteristics when they are bound to a specific ion species; others respond to changes in membrane potential. It is therefore possible to use these indicator fluorochromes to measure important physiological parameters optically, such as free calcium levels, cellular pH and membrane potential. The application of fluorescence microscopy to living cells is often not a straightforward matter. Problems such as low fluorescent intensity and phototoxicity often arise. Given the state of the field and the potential power of these techniques it is timely that the American Society for Cell Biology has devoted Volumes 29 and 30 of their series Methods in Cell Biology to the subject of fluorescence microscopy of living cells in culture. The editors are to be commended for producing an excellent, informative reference work which will be of great use to all who are either using these techniques, or contemplating using them. All the contributors seem to have done their best to help and inform rather than to dazzle and intimidate. There are many hints and tips included that are not present in the original papers. The topics range from basic fluorescence microscopy to methods for evaluating the performance of optical instruments; the incorporation of macromolecules into cells to fluorescent analogue
Methods in Cell Biology, Vols 29 and 30; Fluorescence Microscopy of Living Cells in Culture, Parts A and B edited by Yu-I.i Wang and D. Lansing Taylor, Academic Press, 1989. Part A $59.00 (xi + 333 pages) ISBN 0 12 564129 X Part B $94.00 (xiv + 503 pages) ISBN 0 12 564130 3 Even the most die-hard biochemist or molecular biologist now appreciates that a cell is not simply an oily bag containing a complex mixture of reacting chemicals, but rather it is an intricate, dynamic, selfassembling machine with a rich variety of component parts. Many basic cellular functions such as karyokinesis, cytokinesis, locomotion and secretion operate by the assembly and disassembly of spatially ordered mechanisms. This, in turn, requires the vectored intracellular transport of specific subcellular components. In order to understand how such a machine works, it is essential to have both information on the structure and organization of the component parts, together with a knowledge of how these structures change over time. The electron microscope has given us detailed, highresolution snapshots of cellular components but its humble father, 120
ALISON SMITH Universityof Cambridge,Departmentof Botany, Downing Street, CambridgeCB2 3EA, UK.
TIBS15-MARCH1990
cytochemistry; from low light level video systems to confocal microscopy; from methods for labelling the cell surface to methods for labelling internal structures; from resonance energy transfer microscopy to how to build a fluorescence recovery after photobleaching instrument; from fluorescent indicators of ion concentrations to fluorescent indicators of membrane potential. The only serious omission from these
A primer for the 'new genetics' Genetics, a Molecular Approach by T.A. Brown, Van N o s t r a n d Reinhold, 1 9 8 9 . £ 1 6 . 9 5 (xvii + 3 8 7 pages) ISBN 0 2 7 8 0 0 0 4 1 X
One is tempted when reviewing a new book to be outrageous: either in condemnation or adulation. In this instance, such an outburst would be inappropriate. This book is not packed with new or controversial ideas. The author has attempted to write a basic, simple genetics textbook aimed at students for whom genetics is part of a broader degree course; I believe he has succeeded. As an occasional teacher of the 'new genetics' to the lay public, I do feel that there is a need for this type of textbook. To understand this book requires very little previous knowledge of biology. Anyone could pick it up and read it from cover to cover in a few days and be a lot wiser for it. The book's strongest point is its approachable, almost 'chatty' style. Words are kept to a minimum but without becoming telegraphic. The aim of each chapter is clearly set out and one chapter leads on to the next in a logical order. The reader is continually coaxed into reading on. The first section of the book starts with Mendel and covers the historical aspects of the discovery of the chemical nature of the gene. We then get down to the backbone of molecular biology: DNA structure, transcription, translation and replication. The basic facts and concepts are explained clearly and simply. The stage is set for the rest of the book in which genetic systems are described within the context of the cell or whole organism. This covers the organization of the genome in viruses, bacteria and eukaryotes and this naturally leads on to the basic principles of linkage and recombination. Each chapter ends with a reading list.
volumes is a discussion of the photochemistry of bleaching, information of the bleaching rate of commonly used fluorochromes and suggestions as to how to minimize this phenomenon in practical situations. This is a crucial issue for all who work in this field because the bleaching rate of the fluorochrome is one of the major determinants of the overall detection sensitivity. ! suspect that this is still such a murky field that no one could be persuaded to comment on it. 1 hope
Some of the general references here are good but I do not feel that the person who buys or uses this book is going to get anything out of references to the original work, for example: 'Hoagland et al. (1958)
A Soluble Ribonucleic Acid Intermediate in Protein Synthesis, Journal of Biological Chemistry 231,241-257', as to understand this type of paper requires a great deal of biochemical knowledge. It would have been better to suggest other more detailed textbooks. A set of questions is found at the end of each chapter (answers to the simplest of these can be found at the back of the book). This helps the reader to determine how much he or she has actually understood. It is a shame that some of the figures are poor and with a little more thought these could so easily have been better. The use of computer
Review copies of the following have been received. Books which have been reviewed in full in TIBS are not included.
this omission will stimulate some directed research effort into this important topic. Apart from this, I can thoroughly recommend these volumes to anyone who is interested in the problem of how to view the complex processes that take place in a living cell directly.
JOHN WHITE MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
facsimiles of scientists' portraits is unfortunate. My only other minor criticism is that the authors have defined 'coding strand' in the opposite way to current convention. Presumably this is an e r r o r and can be corrected in the next edition. This book does not replace Lewin's Genes I11,which remains the book of choice for those students or researchers who are actively working, or hoping to work, in this field. For those who are perhaps studying 'Applied Biology' or 'Human Sciences' and need a shorter, more digestible textbook, it is ideal.
E M M A WHITELAW Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
P. M. Hawkey and D. A. Lewis (eds) Medical Bacteriology: A Practical Approach IRL Press, 1989. £38.00 (hbk)/£25.00 (pbk) (xix + 321 pages)ISBN 0 19 963009 7
B. P. Gaber, J. M. Schnur and D. Chapman (eds) Biotechnological Applications of Lipid Microstructures, Vol. 238 Plenum Press, 1989. $95.40 (xi + 397 pages) ISBN 0 306 43014 2
Jacek Hawiger (ed.) Methods in Enzymology, Vol. 169, Part A Academic Press, 1989. $65.00 (xxviii + 512 pages) ISBN 0 12 182070 X
J. W. Gorrod, O. Albano and S. Papa (eds) Molecular Aspects of Human Disease, Vol. 2 John Wiley & Sons, 1989. £39.95 (v + 284 pages) ISBN 0 7458 0587 6
W. Hewitt and S. Vincent Theory and Application of Microbiological Assay Academic Press, 1989. £28.50 (xii + 323 pages) iSBN 0 12 346445 5
H. Greiling and J. E. Scott Keratan Sulphate, Chemistry, Biology, Chemical Pathology Biochemical Society, 1989. £25.00/$48.00 (272 pages)ISBN 0 904498 25 5
W. H. Hori and A. Heiland (eds) Proteases II Potential Role in Health and Disease, Vol. 240 Plenum Press, 1988. $114.00 (xvi + 574 pages) ISBN 0 306 43018 5
J. R. Harris and A. H. Etemadi (eds) Subcellular Biochemistry, Vol. 14, Artificial and Reconstituted Membrane Systems Plenum Press, 1989. $107.40 (xvii + 501 pages) ISBN 0 306 43055 X
S. U. Kim (ed.) Myelination and Demyelination Implications for Multiple Sclerosis Plenum Press, 1989. $65.00 (viii + 275 pages)ISBN 0 306 43118 1
D. E. Hathway Molecular Mechanisms of Herbicide Selectivity Oxford University Press, 1989. £32.50 (x + 214 pages) ISBN 0 19 857 642 0 T. J. Hassold and C. J. Epstein (eds) Molecular and Cytogenetic Studies of Non.Disjunction, Vol. 311 Alan R. Liss, 1989. $72.00 (xiv + 376 pages) ISBN 0 8451 5161 4
V. Kostka (ed.) Protein of Retroviruses Walter de Gruyter, 1989. DM 198.00 (xi + 206 pages) ISBN 0 899255 17 5 A. M. Krstulovic (ed.) Chiral Separations by HPLC Ellis Horwood, 1989. £69.50 (v + 548 pages) ISBN 0 7458 0331 8 121
aspect that is different from most of the earlier books in the series is that it is available in spiralbound format - much more 'practical' for the bench than paperback. The intended audience for Methods in Plant Molecular Biology is quite different. It is specifically designed for use as a manual for an undergraduate laboratory course, and is based on one which is run at the University of Illinois. The experiments described introduce a wide range of different techniques, starting from basic cloning and analysis of DNA, through translation in vitro by isolated chloroplasts and the wheat germ system, and concluding with dideoxynucleotide DNA sequencing and transformation with Agrobacterium. This course is meant to be done two days a week over a 15-week semester, but it is possible to pick out small groups of experiments to illustrate more specific techniques, such as basic cloning or characterization of a genomic DNA clone, which could be done in shorter periods. Each experiment has an introduction which explains the overall
procedure, and the rationale for doing it. The protocols themselves are clearly presented and easy to follow, and include interesting snippets of information, such as why chloroform is often included in a phenol extraction, or why different proteins give different responses with the Lowry test. In addition, there are highlighted warnings to alert the students to potential dangers to themselves, or to their experimental results, and notes for instructors to facilitate the setting up and running of the practicals. Its value, however, is likely to be confined to someone thinking of instituting a practical course in this area, rather than as a research laboratory manual. In the end, whichever book you buy, I bet that within a short period of time you will be developing your own way of doing things - and then you can write your own 'definitive' manual!
Shedding light on living cells
the light microscope, is being used increasingly to study the dynamics of cellular processes. This renaissance of the light microscope has come about partly because of the development of techniques that facilitate the non-invasive observation of live cells. Phase and Nomarski optics convert small refractive index differences within a sample into intensity changes. More recently, electronic contrast enhancement has pushed the limits of what can be detected by Nomarski optics to the extent that individual microtubules with a diameter of one-twentieth the wavelength of visible light may be clearly discerned in favourable circumstances. Powerful though these techniques are, they are undiscriminating in that all structures that have a different refractive index from their immediate environment will be imaged. Stains with specific affinities have long been used as a method of discriminating a structure of interest from a complex background in fixed cells. Fluorescent stains have become increasingly used because of the high detection sensitivities that may be obtained. Immunofluorescence techniques have been developed to the extent that a specific stain may be constructed for practically any cellular component that possesses a unique hapten. In recent years a number of research groups have developed methods for using selective fluorescent staining techniques in living cells. These methods open up the exciting possibility of being able to follow
the fate of a particular cellular component during normal cellular processes. Furthermore, fluorochromes have been developed that alter their fluorescence emission characteristics when they are bound to a specific ion species; others respond to changes in membrane potential. It is therefore possible to use these indicator fluorochromes to measure important physiological parameters optically, such as free calcium levels, cellular pH and membrane potential. The application of fluorescence microscopy to living cells is often not a straightforward matter. Problems such as low fluorescent intensity and phototoxicity often arise. Given the state of the field and the potential power of these techniques it is timely that the American Society for Cell Biology has devoted Volumes 29 and 30 of their series Methods in Cell Biology to the subject of fluorescence microscopy of living cells in culture. The editors are to be commended for producing an excellent, informative reference work which will be of great use to all who are either using these techniques, or contemplating using them. All the contributors seem to have done their best to help and inform rather than to dazzle and intimidate. There are many hints and tips included that are not present in the original papers. The topics range from basic fluorescence microscopy to methods for evaluating the performance of optical instruments; the incorporation of macromolecules into cells to fluorescent analogue
Methods in Cell Biology, Vols 29 and 30; Fluorescence Microscopy of Living Cells in Culture, Parts A and B edited by Yu-I.i Wang and D. Lansing Taylor, Academic Press, 1989. Part A $59.00 (xi + 333 pages) ISBN 0 12 564129 X Part B $94.00 (xiv + 503 pages) ISBN 0 12 564130 3 Even the most die-hard biochemist or molecular biologist now appreciates that a cell is not simply an oily bag containing a complex mixture of reacting chemicals, but rather it is an intricate, dynamic, selfassembling machine with a rich variety of component parts. Many basic cellular functions such as karyokinesis, cytokinesis, locomotion and secretion operate by the assembly and disassembly of spatially ordered mechanisms. This, in turn, requires the vectored intracellular transport of specific subcellular components. In order to understand how such a machine works, it is essential to have both information on the structure and organization of the component parts, together with a knowledge of how these structures change over time. The electron microscope has given us detailed, highresolution snapshots of cellular components but its humble father, 120
ALISON SMITH Universityof Cambridge,Departmentof Botany, Downing Street, CambridgeCB2 3EA, UK.
TIBS15-MARCH1990
cytochemistry; from low light level video systems to confocal microscopy; from methods for labelling the cell surface to methods for labelling internal structures; from resonance energy transfer microscopy to how to build a fluorescence recovery after photobleaching instrument; from fluorescent indicators of ion concentrations to fluorescent indicators of membrane potential. The only serious omission from these
A primer for the 'new genetics' Genetics, a Molecular Approach by T.A. Brown, Van N o s t r a n d Reinhold, 1 9 8 9 . £ 1 6 . 9 5 (xvii + 3 8 7 pages) ISBN 0 2 7 8 0 0 0 4 1 X
One is tempted when reviewing a new book to be outrageous: either in condemnation or adulation. In this instance, such an outburst would be inappropriate. This book is not packed with new or controversial ideas. The author has attempted to write a basic, simple genetics textbook aimed at students for whom genetics is part of a broader degree course; I believe he has succeeded. As an occasional teacher of the 'new genetics' to the lay public, I do feel that there is a need for this type of textbook. To understand this book requires very little previous knowledge of biology. Anyone could pick it up and read it from cover to cover in a few days and be a lot wiser for it. The book's strongest point is its approachable, almost 'chatty' style. Words are kept to a minimum but without becoming telegraphic. The aim of each chapter is clearly set out and one chapter leads on to the next in a logical order. The reader is continually coaxed into reading on. The first section of the book starts with Mendel and covers the historical aspects of the discovery of the chemical nature of the gene. We then get down to the backbone of molecular biology: DNA structure, transcription, translation and replication. The basic facts and concepts are explained clearly and simply. The stage is set for the rest of the book in which genetic systems are described within the context of the cell or whole organism. This covers the organization of the genome in viruses, bacteria and eukaryotes and this naturally leads on to the basic principles of linkage and recombination. Each chapter ends with a reading list.
volumes is a discussion of the photochemistry of bleaching, information of the bleaching rate of commonly used fluorochromes and suggestions as to how to minimize this phenomenon in practical situations. This is a crucial issue for all who work in this field because the bleaching rate of the fluorochrome is one of the major determinants of the overall detection sensitivity. ! suspect that this is still such a murky field that no one could be persuaded to comment on it. 1 hope
Some of the general references here are good but I do not feel that the person who buys or uses this book is going to get anything out of references to the original work, for example: 'Hoagland et al. (1958)
A Soluble Ribonucleic Acid Intermediate in Protein Synthesis, Journal of Biological Chemistry 231,241-257', as to understand this type of paper requires a great deal of biochemical knowledge. It would have been better to suggest other more detailed textbooks. A set of questions is found at the end of each chapter (answers to the simplest of these can be found at the back of the book). This helps the reader to determine how much he or she has actually understood. It is a shame that some of the figures are poor and with a little more thought these could so easily have been better. The use of computer
Review copies of the following have been received. Books which have been reviewed in full in TIBS are not included.
this omission will stimulate some directed research effort into this important topic. Apart from this, I can thoroughly recommend these volumes to anyone who is interested in the problem of how to view the complex processes that take place in a living cell directly.
JOHN WHITE MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
facsimiles of scientists' portraits is unfortunate. My only other minor criticism is that the authors have defined 'coding strand' in the opposite way to current convention. Presumably this is an e r r o r and can be corrected in the next edition. This book does not replace Lewin's Genes I11,which remains the book of choice for those students or researchers who are actively working, or hoping to work, in this field. For those who are perhaps studying 'Applied Biology' or 'Human Sciences' and need a shorter, more digestible textbook, it is ideal.
E M M A WHITELAW Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
P. M. Hawkey and D. A. Lewis (eds) Medical Bacteriology: A Practical Approach IRL Press, 1989. £38.00 (hbk)/£25.00 (pbk) (xix + 321 pages)ISBN 0 19 963009 7
B. P. Gaber, J. M. Schnur and D. Chapman (eds) Biotechnological Applications of Lipid Microstructures, Vol. 238 Plenum Press, 1989. $95.40 (xi + 397 pages) ISBN 0 306 43014 2
Jacek Hawiger (ed.) Methods in Enzymology, Vol. 169, Part A Academic Press, 1989. $65.00 (xxviii + 512 pages) ISBN 0 12 182070 X
J. W. Gorrod, O. Albano and S. Papa (eds) Molecular Aspects of Human Disease, Vol. 2 John Wiley & Sons, 1989. £39.95 (v + 284 pages) ISBN 0 7458 0587 6
W. Hewitt and S. Vincent Theory and Application of Microbiological Assay Academic Press, 1989. £28.50 (xii + 323 pages) iSBN 0 12 346445 5
H. Greiling and J. E. Scott Keratan Sulphate, Chemistry, Biology, Chemical Pathology Biochemical Society, 1989. £25.00/$48.00 (272 pages)ISBN 0 904498 25 5
W. H. Hori and A. Heiland (eds) Proteases II Potential Role in Health and Disease, Vol. 240 Plenum Press, 1988. $114.00 (xvi + 574 pages) ISBN 0 306 43018 5
J. R. Harris and A. H. Etemadi (eds) Subcellular Biochemistry, Vol. 14, Artificial and Reconstituted Membrane Systems Plenum Press, 1989. $107.40 (xvii + 501 pages) ISBN 0 306 43055 X
S. U. Kim (ed.) Myelination and Demyelination Implications for Multiple Sclerosis Plenum Press, 1989. $65.00 (viii + 275 pages)ISBN 0 306 43118 1
D. E. Hathway Molecular Mechanisms of Herbicide Selectivity Oxford University Press, 1989. £32.50 (x + 214 pages) ISBN 0 19 857 642 0 T. J. Hassold and C. J. Epstein (eds) Molecular and Cytogenetic Studies of Non.Disjunction, Vol. 311 Alan R. Liss, 1989. $72.00 (xiv + 376 pages) ISBN 0 8451 5161 4
V. Kostka (ed.) Protein of Retroviruses Walter de Gruyter, 1989. DM 198.00 (xi + 206 pages) ISBN 0 899255 17 5 A. M. Krstulovic (ed.) Chiral Separations by HPLC Ellis Horwood, 1989. £69.50 (v + 548 pages) ISBN 0 7458 0331 8 121