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doi: 10.1242/10.1242/jcs.00257
Book Review |
edited by J. A. Kiernan and I. Mason
Portland Press (2002)
392 pages. ISBN 1-85578-141-7 £110
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Amanda is fictitious, but anyone who has written a textbook recently might recognise her. She is a young, well-dressed publishing person with little knowledge of science and a brisk, menacing manner. As soon as I open this book, I begin to imagine her at work. She is trying to conceal her boredom as the editors outline their detailed synopsis of the future work. When at last they finish, she speaks: "We are worried that the book you describe is aimed at the UK research student/postdoc market, which is quite limited. We want you to broaden it to include the undergraduate sector. Could you reduce the number of references, or even eliminate them? And, you must conform to our format."
This book certainly has format: the pages are square and there is an arty thumbnail index. Is this, perhaps, why it is so expensive? The references are given without titles. It is obvious which chapters the unwilling authors were compelled to write by Amanda; those chapters are perfunctory and have very few references.
The first chapter on optical microscopy appears to have been unhappily
written by Ivor Mason (his good stuff is later). Apart from numerous slips
like spelling Nomarski wrong, referring to a `filament' in a mercury arc and
confusing refractive index with density, the problem is extreme brevity. There
is simply not enough explanation to be useful, and there is only one
reference. The second chapter (by Wingate) is not much better. A false
distinction is made between video and CCD cameras and the `chip size' is
incorrectly defined as the diagonal length of the chip [for the complicated
truth, see Holst (Holst,
1998
)]. Noise is said to consist of defective pixels, dark noise
and readout noise, but there is no explanation of shot noise (irreducible
noise due to photon statistics), which is of overwhelming importance at low
signal levels. Moreover, students will be puzzled to learn that an Airy disk
is a series of rings.
Next, there is a really interesting but eccentric chapter by Greenberg and
Boyde, mainly about stereoscopic microscopy. Taking stereo pairs by alternate
masking of left- and right-halves of the exit pupil of the objective is
dismissed as impracticable, but I can confirm that the method of Osborn et al.
(Osborn et al., 1978) for
high-NA immunofluorescence works perfectly, at almost zero cost! It will be
very difficult for readers to understand the Edge microscopes with no diagram,
and a historian may want to know how they differ from the systems invented by
Heimstädt for Reichert in 1920 and 1923
(Hartley, 1993). Finally, there
is a perplexing claim that the problem of low resolution along the optical
axis is somehow ameliorated by using multispot confocal rather than
single-spot microscopy. Another short chapter by Wingate covers photography
with the microscope, but fails to give guidance on the choice between film and
electronic imaging (Entwistle,
1999).
From then on, the authors warm to their theme, which is to impart useful tips and protocols for research. There is an excellent specialised chapter on fluorescent staining of living cells (Johnson) and Kiernan makes a thorough and scholarly job (with 41 references) of fixation. Allison, on tissue processing and sectioning, is also good, though it would be nice to know why the safe xylene substitutes now used in schools are `inferior'. Kiernan's account of stains is useful, but it is weak on fluorescent stains. Immunohistochemical methods are also well treated by Susan Van Noorden. Mason's account of new variants of the ancient method of staining of the skeleton in whole animals is useful, but misses the modern use of fluorescence in combination with confocal microscopy for small and intricate skeletons. As the chapters get more specialised they improve. This certainly applies to the metabolic mapping chapter by Cornelis Van Noorden and Frederiks and to the concluding chapters by Mason with Irving and Gait.
I would recommend this book for researchers because the protocols are extremely useful. Thanks to Amanda, it is not as specialised and scholarly as it could have been. Her fingerprint is clear on the reference lists, which are often hopelessly short, and because the references are printed without titles, a student cannot see which is a general reference for a more thorough understanding and which is a specialised paper. I suspect that Amanda will now have moved to a different post, where she is in charge of Focus meetings.
MRC Laboratory of Molecular Biology, Cambridge, UK
References
Entwistle, A. (1999). A comparison between the use of a high-resolution CCD camera and 35mm film for obtaining coloured micrographs. J. Microscopy 192, 81-89.
Hartley, W. G. (1993). The Light Microscope: Its Use and Development. Oxford: Senecio Publishing.
Holst, G. C. (1998). CCD arrays, cameras and displays, 2nd edn. Bellingham, WA: SPIE Optical Engineering Press.
Osborn, M., Born, T., Koitzsch, H.-J. and Weber, K. (1978). Stereo immunofluorescence microscopy: 1. Three-dimensional arrangement of microfilaments, microtubules and tonofilaments. Cell 14,477 -488.[CrossRef][Medline]
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