Edited by Robin Harris, John Graham and David Rickwood
Wiley & Sons
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
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.
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.
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.
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.
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.
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.
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.
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.
Division of Genomic Medicine
University of Sheffield