See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/229839060

Plant microtechnique and microscopyBy Steven E. Ruzin. xi+322 pages.

Oxford, UK: Oxford University Press, 1999. £32.50 p/b. ISBN 0 19 508956 1.

Article in New Phytologist · July 2008

DOI: 10.1046/j.1469-8137.2000.00735.x

CITATIONS READS
2 18,315

1 author:

Nigel Chaffey
Bath Spa University
238 PUBLICATIONS 1,524 CITATIONS

SEE PROFILE

All content following this page was uploaded by Nigel Chaffey on 01 July 2018.

The user has requested enhancement of the downloaded file.

REVIEWS New Phytol. (2000), 148, 57–58
Books
Microtechnique
Plant microtechnique and microscopy
By Steven E. Ruzin.
xij322 pages. Oxford, UK : Oxford University
Press, 1999. £32n50 p\b. ISBN 0 19 508956 1.
There is a long and noble tradition of plant microtechnique
texts, including such classics as Johansen (1940), Sass
(1958), Jensen (1962), and O’Brien & McCully (1981).
However, for many years the importance of studying plant
anatomy and structure as a unified discipline (the physio-
logical plant anatomy of Haberlandt (1914)) has been
much neglected. And the ‘ gene revolution ’ of the 1970s
and the subsequent supremacy of molecular-genetical
technologies, threatened to make such ‘ old-fashioned ’
subjects disappear for ever. Fortunately, with the timely
realization that the site of gene action is within the cell,
there is an increased demand for microscopists\anatomists
to place this ‘ molecular work ’ in its proper context. This
has led to something approaching a renaissance in the
practice of plant microtechnique. This modern ‘ molecular
plant anatomy ’ provides the background to the present
book by Ruzin, and some of the justification for its
existence.
The book begins with a chapter entitled ‘ Quick start ’.
This gives an overview of the techniques that are dealt
with in more depth later in the book, and is intended for
use by those who already have some experience in
microtechnique. Following ‘ Microscopy ’, which deals
with basic principles of microscopy (including sections on
confocal and wide-field deconvolution microscopy), there
are chapters devoted to the art and science of micro-
technique itself. Accordingly, we have : ‘ Chemical
fixation’(principles and practice, with welcome sections on
use of the microwave oven) ; ‘ Tissue dehydration ’ (in-
cluding less widely known techniques such as use of rapid
dehydrating agents like dimethoxypropane) ; ‘ Infiltrating
and embedding tissues’ (primarily paraffin, Steedman’s
low-melting-point wax, and acrylic resins) ; ‘ Sectioning
and mounting ’ (including microtome discipline, and
section-adhesion) ; ‘ Staining ’ (most – if not all – the ‘ old
faithfuls ’ are there, for example, safranin, orange G, fast
green, haematoxylin, crystal violet, toluidine blue, meth-
ylene blue, plus useful sections dealing with coverslip
mounting media, and the problematical subject of staining
epoxy resin sections) ; ‘ Alternative methods of microtomy ’
(such as the vibrating and freezing microtomes, which are
currently enjoying a new lease of life with present-day
developmental cell biological techniques) ; ‘ Special
methods ’ (including maceration and tissue-clearing, pre-
serving colour in whole mounts, and repairing broken
slides) ; ‘ Microtechnique notes and problem solving ’ (full
of ‘ tried-and-tested ’ trouble-shooting tips and tech-
niques) ; ‘ Histochemistry and cytochemistry ’ (colour,
contrast and fluorescence techniques for identification of
cell components and enzymes), and ‘ Localization of
molecular targets in tissues ’ (including principles and
practice of immunolocalization, tissue-printing, a detailed
consideration of, and protocol for, in situ hybridization
(sections and whole mounts), autoradiography, and the
TUNEL assay for programmed cell death) ; Appendices
(sections dealing with : toxicity of laboratory chemicals ;
making buffers ; molarities of acids and bases ; the mercury
lamp ; supplier details ; and a detailed consideration of
optics) ; and a Bibliography (over 550 references, the back
cover informs us).
Who is it aimed at ? In the Introduction, Ruzin refers to
‘ students ’. Whether the intention was to target under-
graduates specifically, or to embrace all those who need to
understand such techniques generally, is not explicit. But,
in view of the importance of the subject, and the fact that
it appears to be taught less and less these days, I suggest
that it should be consulted by all who require an
introduction to plant microtechniques, whether just to
understand what their ‘ microtechnical ’ colleagues are
doing, or to encourage them to have a go themselves.
In keeping with the idea that one should use whatever
means are available to further one’s research, it is inevitable
that sources of microtechnique information on the Internet
are mentioned. Even the most cursory glance through the
references (which are an impressive collection) will show
numerous citations of web addresses. As the amount of
available information increases, such sources are likely to
be cited, and used, more and more, probably to the
detriment of ‘ hard copy ’ sources. However, postings to
the Internet are often much abbreviated (and not always
intended for such widespread dissemination). As ephem-
eral electronic texts seem set to replace printed ones, there
is also a danger that the original sources of the data will be
forgotten, and gems of information lost for ever. So I
would like to take this opportunity to set one record
straight in a small way. Whilst I am flattered to see
reference to my posting to the microscopy list-server about
toluidine blue-quenching of fluorescence, in the interests
of fairness, I would like to point out that the source of this
procedure is Smith & McCully (1978).
Unavoidably one will compare this book with its
predecessors. Perhaps the most appropriate comparison is
with O’Brien and McCully. How does Ruzin fare ? In
short, very well. One of the great features of O’Brien and
McCully is the feeling it gives that the authors have
actually tried the techniques and are speaking from a
position of knowledge based on hands-on experience. I get
a similar feeling with Ruzin. The book is written in a
friendly, accessible style (where the abundant lists, tables
and protocols permit ‘ proper writing’). It also contains
some nice touches, for example, the table giving details of
how to remove various stains from hands (appropriately,
in the chapter on staining). Although all the standard plant
techniques appear to be there ; its major strength is its
 58 REVIEWS Books
coverage of new (i.e. post-O’Brien and McCully) tech-
niques, such as confocal microscopy, in situ hybridization,
the widespread use of Steedman Wax and butyl-methyl-
methacrylate for imunoflorescence work, and TUNEL-
staining.
What is wrong with the book ? It is rare to find a single-
authored book these days, particularly one that covers the
enormous scope of botanical microtechnique. Almost
inevitably, such a book will not cover every aspect with
equal thoroughness. For instance, as a student of the
developmental biology of trees, I am disappointed (but not
surprised) that there is little specifically dealing with
microtechnique of this demanding group of plants. In view
of the importance of trees and the great interest in
molecular aspects of wood formation (e.g. Savidge et al.,
2000), I would hope that deficiency will be made good in
future editions, or perhaps it will be left to more specialist
tomes, such as Chaffey (2001). In some respects that is a
minor quibble. Trees are not everyone’s cup of tea, and
practically all the techniques that are included should work
with Arabidopsis (which is probably of more immediate
concern to the molecularly inclined).
However, a more serious deficiency is the absence of
photomicrographs. One is drawn to the book by the three
photomicrographs on the cover (although nowhere could I
find a legend telling me what cell secrets they were
revealing), but, once ensnared, one is given no more. I do
not understand how a book devoted to such a visual subject
as plant microtechnique can be published without any
examples of the results (though I expect we will be told by
the publisher that it is something to do with cost). If we are
to enthuse a new generation and encourage them to make
the voyage of discovery that awaits all who probe the
interior of the cell, I feel that lack of micrographs (which
are so essential to proper understanding and appreciation
of different staining techniques, for example) is an
opportunity missed. Still, at least the book has an index
(which was so conspicuously lacking in O’Brien and
McCully).
For many years there have been rumours of a second
edition of O’Brien and McCully’s book, but none has been
forthcoming, although in the two decades since it was first
published, much has happened in the field of plant
microtechnique. However, now there is no need for
O’Brien and McCully II because we now have Ruzin I.
There is a new demand for plant microtechnical expertise,
and this most useful book fulfils that. It should help a new
generation come to grips with this most important, and
satisfying, branch of botany. Every plant-science labora-
tory should have a copy.
N. J. C
IACR, Long Ashton Research Station, Department of
Agricultural Sciences,
University of Bristol, Long Ashton,
Bristol BS41 9AF, UK
(tel j44 1275 392181 ; fax j44 1275 394281 ;
e-mail nigel.chaffey!bbsrc.ac.uk).

Chaffey NJ, ed. 2001. Wood formation in trees : developmental cell
biology techniques. The Netherlands : Harwood Academic
Publishers. (In press.)
Haberlandt G. 1914. Physiological plant anatomy. London, UK :
Macmillan and Co., Ltd.
View publication stats
Jensen WA. 1962. Botanical histochemistry : principles and
practice. San Francisco, CA, USA : W. H. Freeman and Co.
Johansen DA. 1940. Plant microtechnique, 1st edn. New York,
USA : McGraw-Hill Book Co. Ltd.
O’Brien TP, McCully ME. 1981. The study of plant structure :
principles and selected methods. Melbourne, Vic., Australia :
Termacarphi Pty Ltd.
Sass JE. 1958. Botanical microtechnique, 3rd edn. London, UK :
Constable and Company Ltd.
Savidge R, Barnett J, Napier R, eds. 2000. Cell and molecular
biology of wood formation. Oxford, UK : Bios Scientific
Publishers.
Smith MM, McCully ME. 1978. Enhancing aniline blue
fluorescent staining of cell wall structures. Stain Technology 53 :
79–85.
Toxic plants
Toxic plants : dangerous to humans and
animals
By Jean Bruneton (translated by Caroline K.
Hatton).
xj545 pages. Andover, UK : Intercept Ltd, 1999.
£92n00 h\b. ISBN 1 898298 62 9.
As claimed by the publishers, this book will be of value to
physicians, pharmacists and veterinarians wanting a quick
answer to possible poisoning incidents and has up-to-date
literature references for those wanting to obtain more
detailed information on symptoms and chemistry. The
book covers wild plant species, garden introductions
and some tropical plants grown as houseplants. The
tropical entries are very limited by necessity of space and
seem to be chosen largely for interesting chemistry or
symptoms. In some cases the toxin details are very full for
plants from other parts of the world (though some will be
more common in France), for example alkaloids in
legumes, but in others cases the details are rather sparse
(e.g. oestrogenic compounds of clovers and endophyte
toxins in grasses). The chemical details are presented in a
format useful to chemists and there is also an appendix
giving a summary of phytochemistry for nonexperts.
Throughout the text are descriptions of the plants with
nice black-and-white drawings and chemical structures,
and distribution of the chemicals in parts of the plants are
given when known.
The author gives poisoning statistics for Europe and the
USA and draws such conclusions as : of 598 poisoning
deaths in children under the age of 10 in 20 years in
Britain, only two were associated with plants (and there
was doubt about the association of one of those). There is
a very informative and balanced introduction to the risks
of phytotherapy and consumption of wild foods. The risk
of overdosing with herbal products is discussed, as is the
possibility of side effects which might be difficult to assess.
The dangers of misidentification are pointed out, with
examples. Professor Bruneton rightly points out that the
toxic components of plants are still far from fully known.
In conclusion, this is a very useful book on toxic plants,
giving botanical details of the plants, chemical information
and symptoms. It is also very well referenced.
R J. N
Research Director, MolecularNature Ltd.
Plas Gogerddan, Aberystwyth, SY23 3EB, UK
(tel j 44 1970 823201 ; fax j 44 1970 823209 ;
e-mail robert.nash!bbsrc.ac.uk).