Getting the picture: Video Microscopy ( Methods in Cell Biology, Vol. 56), edited by Greenfield Sluder and David E. Wolf

Abstract

Academic Press, 1998. £67.00 (hbk), £44.95 (pbk) (334 pages)ISBN 0 12 564158 3Video microscopy has profoundly changed the nature of cell biology, introducing new experimental possibilities for examining molecular interactions in vitro and in vivo. Digitally enhanced differential interference contrast (DIC) microscopy has been used to characterize motor proteins in motility assays1xSalmon, E.D. Trends Cell Biol. 1995; 5: 154–158Abstract | Full Text PDF | Scopus (15)See all References1 and push the limits of positional tracking beyond the resolution of the light microscope to the nanometer level2xSheetz, M.P. et al. Nature. 1989; 340: 284–288Crossref | PubMedSee all References2. Alternatively, video cameras more photosensitive than the human eye have made it possible to follow the interactions between fluorescently labelled microtubules and focal contacts in living cells3xKavarina, I., Rottner, K., and Small, J.V. J. Cell Biol. 1998; 142: 181–190Crossref | Scopus (213)See all References3. Initially, the technology was borrowed, for example, from television and the military, but now there exists a market full of imaging products designed solely for biological microscopy. Given the sophistication, to say nothing of the cost, of imaging equipment, careful consideration is required in assessing the imaging tasks at hand and assembling a system of appropriate components, especially when quantitative results are desired. Furthermore, the proper adjustment and use of video and optical equipment is often crucial to the final quality of image data.Video Microscopy is both a clearly written introductory text and a useful reference volume for cell biologists. It begins by introducing video basics and progresses to sophisticated imaging applications. The second chapter, on low-light microscopy, serves to introduce basic characteristics of video systems such as noise, sensitivity, quantum efficiency and dynamic range, and compares different types of camera detectors. This chapter is informative, well written and could be said to serve as a cornerstone for the rest of the volume. Although the discussion is necessarily technical, the language is clear and many examples are provided. A following chapter builds on this introduction with a detailed comparison of the performance of intensified and cooled charge-coupled device (CCD) cameras, two systems that frequently compete for use at the lowest levels of photon detection. Successive chapters cover digital images and their manipulation, beginning with the conversion of an analogue image to digital form by the camera’s CCD chip. Then the hardware components of a digital image processor are introduced and image processing is discussed, beginning with simple contrast enhancement and frame averaging and progressing to more complex transformations and convolutions. The final chapter of the book even manages to explain two common methods of digital deconvolution in simple terms.While the first half of the book is largely a technical introduction, the second half is devoted to practical applications, beginning with the detailed description of two fundamentally different video microscopes. The first is a system for high-resolution video enhanced DIC, the second a multi-mode system for collecting paired fluorescence and phase-contrast or DIC images. The second system is of the type appropriate for low-light fluorescence imaging of live cells, such as those expressing GFP, microinjected with fluorescent proteins or treated with other fluorescent reporters. Computer-controlled peripheral devices such as shutters and filter wheels are introduced, which are required for multi-channel fluorescence time-lapse imaging but generally must be purchased separately from the camera and integrated into the system by the user. The last five chapters of the book present some specialized applications, including ratio imaging and the measurement of intracellular Ca2+ and pH.Throughout the book, the same material is often covered from slightly different perspectives, which, far from being redundant, allows cross referencing and ultimately leads to a deeper understanding of what the terms mean. The editors have understandably avoided a discussion of confocal laser scanning microscopy (CLSM) since this technique has been reviewed well elsewhere4xHandbook of Confocal Microscopy. Pawley, J.B. See all References4; however, it would be helpful if CLSM were compared with video with regard to basic system characteristics. I also find the lack of a chapter summarizing general approaches in low light, live-cell imaging to be an unfortunate omission. Finally, it seems inevitable to compare this volume, Video Microscopy, with the volume of the same name by Inoue and Springer5xVideo Microscopy: The Fundamentals. Inoue, S. and Spring, K.R. CrossrefSee all References5, the latter often being referred to simply as ‘the Bible.’ Although Inoue’s book is an incomparably thorough discussion of the subject, the amount of technical information is formidable. By contrast, Sluder and Wolf’s volume presents a practical overview, introducing technical background and a range of techniques in relatively nontechnical language. It thus represents a tutorial approach, appropriate for those who would like to gain rapid access to the sophisticated techniques of video microscopy.