BSCB Newsletter, Winter 2006

Book Reviews

Cell Biology Protocols
Edited by Robin Harris, John Graham and David Rickwood
John Wiley & Sons
ISBN: 0-470-84758-1
January 2006

Cell biology is one of the fundamental areas of research that incorporates a broad range of laboratory methods and techniques. The present book is a collection of a wide variety of such protocols that could be applied by anyone who works on cell biology, from the student to the experienced scientist.

Overall, the format of this book is very helpful, with tables, figures and graphs cleverly supplied where more information is needed. Reagents and materials are described in detail and the steps of the protocols are presented in bullet-points in short but clear sentences that would be easy to follow in praxis. Also, each chapter has a short introduction that orients the reader to the content of the protocols that are to follow.

In specific terms, the book covers six areas of cell biology methods. The first two chapters give a thorough insight into light and electron microscopy. The basic principles of the function of the light and electron microscope are well reviewed and accompanied by detailed yet clear to understand schematic figures. This theoretical explanation is followed by a number of protocols on the processing of biological specimens for microscopy, whether they are tissue sections, cells or organelles.

The next chapter describes methods for primary as well as secondary culture. Starting with the extraction of the cells of interest from fresh tissue, the procedure is explained step-by-step to the method of cell isolation from a heterogeneous culture to cell sub-culture as well as thawing and freezing of cell lines. Cell counting, quantification of cell viability and purification methods are included. Admittedly, there is a degree of variation in cell culturing methods depending on the cell type, however the authors present a very good baseline of protocols with great efficacy regarding any cell type.

The subsequent chapter deals with the purification of subcellular membranes, organelles and organelle components. For example, protocols describe isolation or preparation of nuclei, nucleolei and nuclear membranes, chromosomes, mitochondria and lysosomes as well as separation of the smooth and the rough ER. Some protocols cover the preparation of organelle components of plant tissues. Also a number of assays for quantification of enzymatic activity can be found.

The next chapter is dedicated to the separation of subcellular domains to be used in studies related to membrane trafficking and cell signalling, using different gradient media. Sucrose, Nycodenz and iodixanol gradients are mainly used in the protocols provided in order to fractionate subcellular compartments such as lipid rafts, the apical and basolateral domains of polarised mammalian cells and the ER and Golgi systems.

The last chapter presents a wide variety of in vitro assays of reconstitution in cell biology dealing with nuclear components, cell membrane systems as well as cytoskeletal and fibrillar systems. A few examples of the protocols presented are DNA labelling techniques for cell functional studies, techniques for the study of nuclear–matrix interactions and the uncovering of nuclear matrix for microscopical observation as well as nuclear assembly techniques.

The concept of the use of nanocapsules to improve drug delivery intracellularly is well interpreted, as are apoptosis detection assays and a microarray-based protocol for studies on membrane transport processes. In addition, protocols relating to fibrillar systems, amyloid-b-enzyme interaction and amyloid-b phosphorylation can be found. The final additional chapter contains useful information on chemical safety and the procedure of centrifugation.

Numerous references are provided at the end of each protocol for further reading as well as notes, explanatory comments and tips which often prove to be crucial for the success of a laboratory technique. A valuable possession for every cell biologist's library.

Mary Michailidou
Division of Genomic Medicine
University of Sheffield
M.Michailidou@sheffield.ac.uk