edited by Ammasi Periasamy Oxford University Press (2001) 434 pages. ISBN 1-19-513936-4£75
Published under the auspices of the American Physiological Society, with contributions from many of the luminaries in cell imaging, this book promises much. The subject is hot, but, at first sight, the volume is unappealing. The dust jacket design is crude and garish; the colour figures are all bunched together for cheapness of production. The first few chapters are not outstanding. For example, in the introductory discussion of fluorescence,oxygen, which everyone knows is a key player in phototoxicity, is mentioned merely as a possible fluorescence quencher. Brightness of a fluorophore is equated with a high quantum yield, without mention of the need also for a high extinction coefficient. The pioneering work of Carlsson and Mossberg on the suppression of interchannel cross-talk by the use of pulsed lasers and detectors is incorrectly described as a software innovation. Incredibly,Minsky is the citation for the development of laser scanning microscopy(Minsky, 1957). A more serious error is the explanation of differential interference contrast, which is short, snappy and wrong. Only Spring writes like an experienced teacher, and his material is more fully explained in his previous writings.
At this point, I began to wonder whether the book would represent any advance over existing standard works, such as Inoué and Spring, Sluder and Wolf, and Allan (Inoué and Spring, 1997; Sluder and Wolf,1998; Allan, 1999). Fortunately, the quality rises later. Duchen et al. give useful detail about the imaging of mitochondria and Verkman,Vetrivel and Haggie provide a clear review of fluorescence recovery after photobleaching (FRAP). However, the image processing chapter is, in my opinion, too elementary for the average modern research student, though it might benefit some professors.
Next the subject of multiphoton excitation was tackled. There is an authoritative, introductory, theoretical chapter by So et al., followed by an ostensibly practical one by Diaspro. The latter author turns out to be slightly eccentric. He airily dismisses commercial multiphoton systems as`immature' and advocates the do-it-yourself approach. But his DIY guide is distinctly lacking in practical detail, omitting all mention of YLF lasers or laser safety. Wallace et al. provide an important review of two-photon microscopy in highly scattering real specimens and charmingly named`phantoms'. There is a fine review of embryological applications by Dickinson and Fraser, including work in progress with a biological test system for new dyes with large multiphoton cross-sections.
The best parts of the remainder of the book are concerned with methods that are in transit from physics and have not yet quite arrived in biology. Zipfel and Webb give a physicists' account of the way that both FRAP and fluorescence correlation spectroscopy can benefit from multiphoton excitation, but there is no biologist to explain the significance of the diffusion constants so obtained. Solid, useful reviews follow by König (radiation effects on cells), Herman et al. (fluorescence resonance energy transfer, FRET) and Bastiaens (fluorescence lifetime measurements in the frequency domain), and there is also good teaching by Periasamy (FRET) and Gerritson and Grauw. Enrico Gratton and his associates dazzle us with news from the frontiers of time-resolved imaging spectroscopy. They show how a lab equipped merely with a microscope, two mode-lockable lasers and years of experimental expertise can achieve images with not only high axial resolution but also lifetime information with unprecedented precision without a time-gated detector.
The remaining chapters deal with simpler technologies. Farkas reviews spectral imaging clearly, but, with the commercial release of spectral analysis software, one might wish for more on the algorithms for working out the individual contributions of multiple dyes from the spectral envelope. Axelrod provides a clear account of total internal reflection microscopy(TIRF), but the same figures are slightly better reproduced in his paper listed below (Axelrod, 2001). The basics of laser traps are well explained by Guilford, and there is an update on bioluminescence by Geusz, revealing that aequorin and luciferase have now joined GFP in being transfected into cells. Crucially, however,bioluminescence cannot be imaged effectively by either confocal or multiphoton methods. Finally, a paper on atomic force microscopy by Gad and Ikai shows the possibilities, but also the extreme difficulties, of this instrument.
My general impression is that there are gems here but embedded in a poor matrix, suggesting little or no editing. This is no surprise: what young editor would dare to tackle a luminary at the peak of his or her career to demand better exposition or breadth? An age that values nothing but primary publications gets the review volumes it deserves.