Basic Confocal Microscopy. Edited by Robert L. Price and W. Gray (Jay) Jerome. New York, Springer, 2011. 313 pages. ISBN 978-0-38778-174-7

Flip through the pages of any life sciences journal today and you are likely to find one or more confocal microscopy images presented in the printed research articles. This is because the modern confocal microscope is no longer just a machine that produces “pretty pictures” of a microscopic sample. Indeed, it is truly an analytical instrument, capable of producing massive amounts of quantitative, multidimensional data ~space, time, color, etc.! , which allow us to infer cellularand molecular-scale structure, function, and activities with high specificity and spatio-temporal resolution. It is imperative that new users of the confocal microscope understand every component of the instrument so that they can make the most of its offered capabilities and produce reliable and reproducible data that furthers the science or biological phenomenon they are trying to study. In addition, these new users need to learn about the limitations of the confocal microscope they are working with to avoid producing image artifacts that confuse data interpretation. The “garbage in – garbage out” principle isn’t just for computing—it equally applies to the process of microscopic imaging as well. There exists a need, therefore, for concise teaching aids that encourage new confocalists to completely comprehend the inner workings of the microscope. This new introductory level book attempts to address this very issue. Price and Jerome’s text opens with a couple of chapters that break down and explain the parts that constitute a confocal microscope. A few more chapters are devoted to the basics of fluorescence, digital image capture and processing, and resolution. These concepts help the new user get up to speed on how to design and execute a proper imaging experiment. Much of the information presented in these chapters is already contained in other classic confocal microscopy monographs, such as Alan Hibbs’ Confocal Microscopy for Biologists ~Hibbs, 2004! , albeit in a slightly less condensed form. Basic Confocal Microscopy includes a few more up-to-date descriptions of the newer confocal hardware features and fluorescent probes that were not around when Hibbs’ book was written, however. A few other noteworthy chapters of this new book set it apart from other publications. Chapters 4 and 5 deal with the various methods of preparing fixed or live specimens, as well as immunofluorescence for confocal imaging, something that I feel has been missing from the current confocal microscopy book literature at this level of detail. These chapters provide very clear descriptions of the advantages and disadvantages of the various specimen fixatives that can be purchased commercially or made in-house, the different types of imaging chambers that facilitate live-cell imaging, and also the necessary considerations for antibody labeling. Chapter 9’s step-by-step guide on a typical imaging session alone makes this book a worthy addition to your microscopy library collection because it contains the closest thing the imaging community is likely to ever get in the form of a generic operation manual for a confocal microscope, with full color figures that direct the reader through the windows of the confocal computer graphical user interface. Although the given instructions are for a Zeiss LSM 510 META, they are equally applicable to any type of laser-scanning confocal instrument available on the market today. The authors make quite an effort to explain all of the typical operating parameters and how they can be adjusted and optimized to achieve the best possible imaging results. I truly believe that one could use this chapter as a teachyourself guide for confocal imaging because it is packed with practical information and presented in an easy-tofollow manner. I was also happy to see a short final chapter devoted to the topic of digital imaging ethics, a topic that deserves some attention with new students and researchers, especially given the growing number of cases of image fraud ~unintentional and intentional! that can lead to years of wasted research efforts and funding, as well as public mistrust of scientists ~Rossner & Yamada, 2004; Rossner, 2006! An important reference to Douglas Cromey’s 2010 article on the appropriate use and manipulation of scientific images was noticeably missing, most likely because both publications were in press around the same time ~Cromey, 2010! I mention this article here to the reader for completion’s sake. The do’s and don’ts of proper image archival and preservation are also touched upon in this chapter. Keep in mind that this book is an introductory level text. For the well-seasoned confocalist, I would still endorse other advanced reference books such as Jim Pawley’s Handbook of Biological Confocal Microscopy, which thoroughly explain the underlying theory behind more complex aspects of confocal technology ~Pawley, 2006! I think the core-facility manager could find many uses for Price and Jerome’s book. At 313 pages, this is the kind of text you could hand off to your new facility users and have them read it in a couple of weeks before or during their training sessions, thus saving time and providing the manager with more opportunities to teach new trainees the practical aspects and rules of imaging in their facility with their particular brand of confocal microscope. Overall, I feel the authors have met their goal of making new confocalists aware of the potential of the instruBook Review MicroscopyAND Microanalysis