Seeing neurons at work: Imaging Neurons edited by Rafael Yuste, Frederick Lanni and Arthur Konnerth

Abstract

CSHL Press, 2000. $120.00 (838 pages)ISBN 0 87969 541 2One of the more recently acquired tools of the neuroscientist is the ability to see what’s going on inside living neurons. Ion fluxes previously visualized only on oscilloscopes can now be viewed with great clarity in real time. Similarly, morphological features not previously visible are now readily distinguishable and easily quantified. For a non-physicist like myself, several books have been invaluable resources in my efforts to visualize neuronal calcium dynamics. Books edited by Pawley1xHandbook of Biological Confocal Microscopy. Pawley, J.B. See all References1, Paddock2xConfocal Microscopy Methods and Protocols. Paddock, S.W. See all References2 and Mason3xFluorescent and Luminescent Probes for Biological Activity. Mason, W.T. See all References3 have served as excellent guides in the general practice of cellular imaging, whereas Lambert’s Calcium Signaling Protocols4xCalcium Signaling Protocols. Lambert, D.G. CrossrefSee all References4 has been very helpful in my more specific studies of neurotoxicity-induced changes in intraneuronal calcium concentrations. Imaging Neurons: A Laboratory Manual presents a compilation of a wide range of imaging techniques applied successfully to studies specifically involving neurons. This manual provides a very important addition to the library of anyone whose work, either directly or indirectly, involves neuronal imaging.Authors of the individual chapters in this book have been drawn from a group of people who have taught sections of the Cold Spring Harbor Laboratory summer course ‘Imaging Structure and Function of the Nervous System’. Offered annually since 1991, the course is available to students and researchers interested in or already working in the field of neuronal imaging. As stated in the Preface, the book is intended to be a manual that can be consulted ‘at the bench’ by experimental scientists.There are seven sections in Imaging Neurons, and these are organized in a logical progression from the first on the subject of basic microscopy to later sections detailing specific techniques. Each section begins with a chapter or two covering background information and relevant basic principles. The introductory chapters are followed by the presentation of specific applications in an easy-to-read, well-illustrated format.Section 1 on ‘Microscopes, image acquisition, and the basics of fluorescent imaging’ includes ten chapters. Particularly interesting to me are Chapter 7 on ‘Infrared video microscopy’ and Chapter 10 on ‘Maintaining live cells and tissue slices in the imaging setup’. Both offer very useful information to anyone interested in visualizing neurons within slices of brain tissue.Section 2 deals with the general subject of ‘Confocal microscopy’. The six relatively brief chapters in this section include a succinct and very well-written introduction, two chapters devoted to confocal studies in vivo (Chapters 13 and 14) and two on subcellular Ca2+ ion dynamics (Chapters 15 and 16).Section 3 covers aspects of the more recently developed technique of ‘Multiphoton microscopy’. Three chapters in this section cover the basics of this emerging technology and four chapters describe specific applications. Although I do not use a multiphoton system (and do not foresee being able to use one in the near future), I find the methodology fascinating and the possibility of one day being able to do some of these things intriguing.Photoactivation is covered in Section 4. The most lucid explanation of this process I have ever read is presented in Chapter 24. Other chapters in this section describe studies conducted with caged calcium (Chapter 26) and with caged glutamate (Chapter 27).I bought this book when I saw it at the annual meeting of the Society for Neuroscience, largely because of what I found in Section 5 – ‘Calcium imaging’. Each of the fifteen chapters in this largest section seemed to be directly relevant to work I am conducting or plan to conduct. The first three chapters provide an excellent summary of what is known about fluorescent calcium indicators; five chapters describe calcium imaging in slices; one details methodology for imaging calcium in mitochondria (Chapter 39); and one provides guidelines for assessing calcium activity in developing mammalian retina (Chapter 41).Section 6 deals with ‘Imaging and other aspects of neuronal function’ and includes chapters describing dye-based studies of membrane potential (Chapters 49–51), sodium (Chapter 52) and synaptic vesicle (Chapter 53) dynamics.Section 7 describes the very recent development and use of ‘Genetically engineered fluorescent probes’. Topics covered here include imaging with green-fluorescent protein and luminescence – research areas that are ‘bridging the gap between molecular biologists and physiologists’.The book concludes with some very useful and practical appendices, especially Appendix 3 (Microscopy: lenses, filters, and emission/excitation spectra) and Appendices 5A (Sample reagents) and 5B (Sample protocols for brain slice preparation and neuronal culture).I highly recommend Imaging Neurons be added to the laboratory collection of any researcher interested in, or conducting experiments involving, neuronal imaging.