Archives of Pathology and Laboratory Medicine: Vol. 123, No. 9, pp. 857–858.
Confocal Microscopy, Methods and Protocols
Edited by Stephen W. Paddock, 446 pp, with illus, $99.50, Totowa, NJ, Humana Press, 1999
Laser scanning confocal microscopy has become a powerful tool in cell biology because of its enhanced abilities to image fluorescent markers in both dead and living cells at high resolution in 3-dimensional data sets. The rapid pace of technological advances, particularly in computer interfaces, has brought confocal microscopy into the mainstream of biomedical research. Moreover, the user-friendliness of the newer instruments minimizes the training needed before novices can generate acceptable data from the microscope.
This book, as part of the series Methods in Molecular Biology, J. M. Walker, editor aims “to take the researcher from the bench top, through the imaging process, to the journal page.” Toward that end, 24 contributions (chapters) from confocal microscopists around the world emphasize various biological applications from preparation of different tissues, to 3-dimensional analysis, to presentation of images for publication. Technical details concerning the microscope and its components were intentionally covered only superficially. The editor's decision to keep the technical jargon to a minimum makes for easy reading and will broaden the book's appeal to biologists in many different fields.
The first 3 chapters provide a brief introduction to confocal imaging, some practical considerations for collecting images, and tips on the selection and use of fluorescent probes. Some highlights included a listing of Web sites for additional information, methods for testing resolution, and a fairly complete list of problems and solutions related to the use of fluorochromes, which is an area of rapid commercial development.
Chapters 4 through 11 describe specific applications of confocal microscopy primarily on fixed tissues. The chapters include techniques for double and triple labeling of gene products, confocal microscopy of plant cells, yeast, sea urchin eggs, and frog embryos. The chapter on fluorescent in situ hybridization in Drosophila was particularly interesting. Each chapter followed a similar format, providing information on the equipment and reagents needed for the experiment and detailed, step-by-