Mary Jones, Richard Fosbery,

Jennifer Gregory and Dennis Taylor

Cambridge International AS and A Level

Biology
Coursebook
Third edition
C A MB R I D G E UNI V E R S I T Y P R E S S
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 Contents

Introduction vi 6 Genetic control 103

The structure of DNA and RNA 103
DNA replication 105
1 Cell structure 1 Genes and mutations 109
Why cells? 2 DNA, RNA and protein synthesis 109
Cell biology and microscopy 2 End-of-chapter questions 115
Animal and plant cells have features in common 4
Differences between animal and plant cells 5
Units of measurement in cell studies 6 7 Transport in multicellular plants 118
Electron microscopes 8
Ultrastructure of an animal cell 13 The need for transport systems in multicellular
Structures and functions of organelles 13 organisms 118
Ultrastructure of a plant cell 18 The transport of water 120
Two fundamentally different types of cell 18 Transport in multicellular plants 120
Tissues and organs 20 Translocation 133
End-of-chapter questions 24 Differences between sieve tubes and xylem vessels 137
End-of-chapter questions 138

2 Biological molecules 29
8 The mammalian transport system 144
The building blocks of life 30
Monomers, polymers and macromolecules 30 The mammalian cardiovascular system 144
Carbohydrates 30 Blood plasma and tissue fluid 150
Lipids 37 Lymph 150
Proteins 39 Blood 151
Water 47 Haemoglobin 154
End-of-chapter questions 49 Problems with oxygen transport 157
End-of-chapter questions 160

3 Enzymes 54
9 The mammalian heart 164
Enzymes reduce activation energy 57
The course of a reaction 57 The cardiac cycle 166
Enzyme inhibitors 62 Control of the heart beat 168
End-of-chapter questions 63 End-of-chapter questions 170

4 Cell membranes and transport 69 10 Gas exchange 174

Phospholipids 69 Lungs 174
Structure of membranes 70 Trachea, bronchi and bronchioles 174
Transport across the cell surface membrane 73 Alveoli 177
End-of-chapter questions 81 End-of-chapter questions 178

5 Cell and nuclear division 86 11 Smoking 182

The nucleus contains chromosomes 86 Tobacco smoke 182
The structure of chromosomes 88 Lung diseases 182
Two types of nuclear division 89 Cardiovascular diseases 185
Mitosis in an animal cell 90 Proving the links between smoking and lung disease 188
Cancer 93 Prevention and cure of coronary heart disease 192
End-of-chapter questions 99 End-of-chapter questions 194

Contents iii
 12 Infectious diseases 197 17 Photosynthesis 287
Worldwide importance of infectious diseases 197 An energy transfer process 287
Cholera 198 The light-dependent reactions of photosynthesis 288
Malaria 200 The light-independent reactions of photosynthesis 290
Acquired immune deficiency syndrome (AIDS) 203 Leaf structure and function 290
Tuberculosis (TB) 208 Chloroplast structure and function 293
Antibiotics 211 Factors necessary for photosynthesis 294
End-of-chapter questions 213 Trapping light energy 295
End-of-chapter questions 297

13 Immunity 217
Defence against disease 217
18 Regulation and control 301
Cells of the immune system 218 Homeostasis 302
Active and passive immunity 225 Excretion 302
Vaccination 226 The structure of the kidney 303
Problems with vaccines 227 Control of water content 311
The eradication of smallpox 228 Nervous communication 314
Measles 230 Hormonal communication 329
End-of-chapter questions 231 Plant growth regulators 336
Electrical communication in plants 339
End-of-chapter questions 340
14 Ecology 235
Energy flow through organisms and ecosystems 236
Matter recycling in ecosystems 241
19 Inherited change 347
The nitrogen cycle 241 Meiosis 347
End-of-chapter questions 245 Genetics 348
Genotype affects phenotype 351
Inheriting genes 352
15 Advanced practical skills 248 Multiple alleles 354
Sex inheritance 355
Experiments 248
Sex linkage 355
Variables and making measurements 249
Dihybrid crosses 357
Estimating uncertainty in measurement 257
The 2 (chi-squared) test 359
Recording quantitative results 258
Mutations 361
Constructing a line graph 259
Environment and phenotype 363
Constructing bar charts and histograms 260
End-of-chapter questions 363
Drawing conclusions 261
Describing data 261
Making calculations from data 261
Explaining your results 263
20 Selection and evolution 367
Identifying sources of error and suggesting Natural selection 368
improvements 263 Evolution 370
Drawings 264 The Darwin–Wallace theory of evolution by natural
End-of-chapter questions 265 selection 374
Species and speciation 375
Artificial selection 377
16 Energy and respiration 268 End-of-chapter questions 378
The need for energy in living organisms 268
Work 268 21 Biodiversity and conservation 382
ATP 270
Respiration 273 The five-kingdom classification 382
Anaerobic respiration 279 Maintaining biodiversity 385
Respiratory substrates 280 Endangered species 385
End-of-chapter questions 283 End-of-chapter questions 392

iv Contents
22 Gene technology 395 26 Planning, analysis and evaluation 468
Gene technology 395 Planning an investigation 468
Benefits of gene technology 399 Analysis, conclusions and evaluation 473
Potential hazards of gene technology 400 End-of-chapter questions 480
Social and ethical implications of genetic engineering 402
Electrophoresis 403
Cystic fibrosis 404
The genetic counsellor 407 Appendix 1 Amino acid R groups 484
Genetic screening 409
End-of-chapter questions 411
Appendix 2 DNA triplet codes 485
23 Biotechnology 414 Glossary 486
Mining with microorganisms 414
Large-scale production techniques 416
Advantages of batch and continuous culture 419 Index 501
How penicillin works 420
Immobilising enzymes 422
Monoclonal antibodies 424
Acknowledgements 510
End-of-chapter questions 427

CD-ROM
24 Crop plants 431
Cereal crops 431 Advice on how to revise for
Maize 433 and approach examinations
C4 plants 434
Adaptations for difficult environments 437
Crop improvement 440 Chapter summaries
End-of-chapter questions 447

Multiple choice tests for

25 Aspects of human reproduction 451 Chapters 1–14
Gametogenesis 453
Human menstrual cycle 456 Answers to self-assessment questions
Birth control 457
Infertility 459
End-of-chapter questions 464 Answers to end-of-chapter questions

Contents v
 Introduction

This new edition is fully updated for the 2014 syllabus Level Syllabus section Chapter
to help you do well in your Cambridge International P Selection and evolution 20
Examinations AS and A level Biology (9700) courses. Applications Q Biodiversity and 21
The book and its accompanying CD-ROM provide a of Biology conservation
self-contained resource for studying these courses, with R Gene technology 22
improved focus on exam preparation. S Biotechnology 23
T Crop plants 24
• Chapters 1–15 provide complete coverage of the
U Aspects of human reproduction 25
AS level syllabus. This is also the first year of study
Planning, analysis and evaluation 26
for A level. The AS syllabus is designed for students
with O level or IGCSE Biology.
There are two new chapters covering practical skills: Chapter 15
• Chapters 16–26 cover all the material for the second
(AS level) and Chapter 26 (A level). Interesting information
year of study for A level. This includes the relevant
that is not required by the syllabus but will aid understanding,
Core material and the Applications of Biology section.
is marked as ‘extension material’ by orange dotted bars.
Extension material is clearly marked with the following Each chapter contains self-assessment questions (SAQs).
symbol E and a dotted line runs down alongside the text These are to help you think about, understand and
to mark this additional content. remember what you have just read. Each chapter ends
Important features of this new edition include the following. with a set of chapter-related questions, ranging from
The sequence of chapters mirrors the sequence of topics formative questions (requiring simple recall or reference to
in the syllabus, which makes it easy to navigate. (Your the text) to more challenging structured or essay questions
teacher may, however, tackle subjects in a different order.) requiring understanding as well as the other skills tested in
Syllabus sections G and H are split into three and two examinations. Some of the questions are past Cambridge
chapters respectively for additional convenience (see table). examination questions so you can familiarise yourself with
the style of the examination questions.
Level Syllabus section Chapter Biology involves many technical terms. Each time a new
AS level A Cell structure 1 term is introduced, it is shown in bold orange and its
Core syllabus meaning explained. The glossary contains definitions of the
B Biological molecules 2 key terms used in the book.
C Enzymes 3 At the end of your course, you will be tested on three sets
D Cell membranes and transport 4 of Assessment Objectives.
E Cell and nuclear division 5
F Genetic control 6 • Knowledge with understanding. You are expected to
G Transport know and understand all the facts and concepts listed
Transport in multicellular plants 7 in the syllabus. These are all covered in this book.
The mammalian transport system
The mammalian heart
8
9
• Handling information and solving problems. Questions
testing these skills expect you to use your knowledge
H Gas exchange and smoking and understanding in an unfamiliar context. A good
Gas exchange 10 knowledge and understanding of this book will enable
Smoking 11
you to approach new situations with confidence.
I Infectious disease 12
J Immunity 13 • Experimental skills and investigations. This
involves practical work. An examination will test
K Ecology 14
your practical skills so try to do plenty of practical
Advanced practical skills 15
A level L Energy and respiration 16
work. Key information is provided on some practical
M Photosynthesis 17 aspects of the course in Chapters 15 and 26.
N Regulation and control 18
O Inherited change 19 Additional help and guidance are available on the
accompanying CD-ROM.

vi Introduction
 1 Cell structure
By the end of this chapter you should be able to:
describe and interpret drawings and photographs the chloroplasts, cell wall, large permanent vacuole,
of typical animal and plant cells as seen using tonoplast and plasmodesmata of plant cells
the light microscope and make microscopical outline the functions of the structures listed above
measurements using an eyepiece graticule and
stage micrometer compare the structure of typical animal and plant cells

be familiar with the units used in cell studies
explain the meanings of, and distinguish between,
the terms resolution and magnification
describe and interpret drawings and photographs
of typical animal and plant cells as seen using the
electron microscope, recognising rough and smooth
endoplasmic reticulum (ER), Golgi apparatus,
mitochondria, ribosomes, lysosomes, cell surface
membrane, centrioles, nucleus (including the nuclear
envelope and nucleolus) and microvilli, as well as
calculate the linear magnification of, and the actual
sizes of, specimens from drawings and photographs
describe the structure of a prokaryotic cell, and
compare and contrast the structure of prokaryotic
cells with that of eukaryotic cells
explain how eukaryotic cells may be organised into
tissues and organs, with reference to transverse
sections of stems, roots and leaves
draw and label low-power plan diagrams of
tissues and organs.

In the early days of microscopy an English scientist, Robert added Virchow’s theory of 1855 that all cells arise from
Hooke, decided to examine thin slices of plant material. pre-existing cells by cell division.
He chose cork as one of his examples. Looking down the
microscope he was struck by the regular appearance of
the structure, and in 1665 he wrote a book containing the
diagram shown in Figure 1.1.
If you examine the diagram you will see the ‘pore-like’
regular structures that Hooke called ‘cells’. Each cell
appeared to be an empty box surrounded by a wall. Hooke
had discovered and described, without realising it, the
fundamental unit of all living things.
Although we now know that the cells of cork are dead,
further observations of cells in living materials were
made by Hooke and other scientists. However, it was
not until almost 200 years later that a general cell theory
emerged from the work of two German scientists. In 1838
Schleiden, a botanist, suggested that all plants are made of
cells, and a year later Schwann, a zoologist, suggested the
same for animals. The cell theory states that the basic unit
of structure and function of all living organisms is the
cell. Now, over 170 years later, this idea is one of the most
familiar and important theories in biology. To it has been Figure 1.1 Drawing of cork cells published by Robert Hooke in 1665.

1 Cell structure 1
 Why cells? Eyepiece lens magnifies
A cell can be thought of as a bag in which the chemistry eyepiece
and focuses the image
of life is allowed to occur, partially separated from the from the objective onto
the eye.
environment outside the cell. The thin membrane which
surrounds all cells is essential in controlling exchange light beam
between the cell and its environment. It is a very effective
barrier, but also allows a controlled traffic of materials across
it in both directions. The membrane is therefore described
as partially permeable. If it were freely permeable, life could
not exist, because the chemicals of the cell would simply objective
Objective lens collects
mix with the surrounding chemicals by diffusion, (page 73). light passing through the
cover slip
specimen and produces a
Cell biology and microscopy glass slide magnified image.

The study of cells has given rise to an important branch of

Condenser lens focuses
biology known as cell biology. Cells can now be studied condenser the light onto the
by many different methods, but scientists began simply specimen held between
by looking at them, using various types of microscope. iris diaphragm the cover slip and slide.
There are two fundamentally different types of Condenser iris
microscope now in use: the light microscope and the light source
diaphragm is closed
electron microscope. Both use a form of radiation in order pathway of light
slightly to produce a
to create an image of the specimen being examined. The narrow beam of light.

light microscope uses light as a source of radiation, while

Figure 1.2 How the light microscope works.
the electron microscope uses electrons, for reasons which
are discussed later.

Light microscopy Golgi apparatus small structures that

The ‘golden age’ of light microscopy could be said to are difficult to identify
cytoplasm
be the 19th century. Microscopes had been available
since the beginning of the 17th century but, when mitochondria
dramatic improvements were made in the quality of
cell surface membrane
glass lenses in the early 19th century, interest among
scientists became widespread. The fascination of
the microscopic world that opened up in biology
inspired rapid progress both in microscope design
and, equally importantly, in preparing material
nuclear envelope
for examination with microscopes. This branch of
chromatin –
biology is known as cytology. Figure 1.2 shows how
deeply staining
the light microscope works. and thread-like nucleus
By 1900, all the structures shown in Figures
nucleolus –
1.3, 1.4 and 1.5, except lysosomes, had been deeply staining
discovered. Figure 1.3 shows the structure of
centriole – always found near nucleus,
a generalised animal cell and Figure 1.5 the has a role in nuclear division
structure of a generalised plant cell as seen with
a light microscope. (A generalised cell shows all Figure 1.3 Structure of a generalised animal cell (diameter about 20 m) as seen
the structures that are typically found in a cell.) with a very high quality light microscope.

2
2 1 Cell structure
 tonoplast – membrane middle lamella – thin layer
surrounding vacuole holding cells together,
contains calcium pectate
cell surface membrane
(pressed against cell wall) plasmodesma –
connects cytoplasm
of neighbouring cells
vacuole – large cell wall of
with central position neighbouring
cytoplasm cell

mitochondria cell wall

nucleolus – chloroplast
deeply staining
grana just visible
Figure 1.4 Cells from the lining of the human cheek ( 500), each nuclear envelope
showing a centrally placed nucleus which is a typical animal cell nucleus
small structures that
characteristic. The cells are part of a tissue known as squamous chromatin –
are difficult to identify
(flattened) epithelium. deeply staining
and thread-like Golgi apparatus

Figure 1.4 shows some actual human cells and Figure 1.6

Figure 1.5 Structure of a generalised plant cell (diameter about 40 m) as
shows an actual plant cell taken from a leaf.
seen with a very high quality light microscope.

SAQ 1.1
Using Figures 1.3 and 1.5, name the structures that
animal and plant cells have in common, those found in
only plant cells, and those found only in animal cells.

Figure 1.6 Photomicrograph of a cell in a moss leaf ( 1400).

1 Cell structure 3
 Box 1A Biological drawing
You need the following equipment: • arrange label lines neatly and ensure they don’t cross
over each other
• pencil (HB)
• annotate your drawing if necessary (i.e. provide
• pencil sharpener
short notes with one or more of the labels in order
• eraser
to describe or explain features of biological interest)
• ruler
• add a scale line at the bottom of the drawing if
• plain paper.
appropriate
Here are some guidelines for the quality of your
drawing:
• use a pencil, not a pen.
An example of a drawing of a section through the stem
• always use a pencil, not a pen of Helianthus is shown below. Biological drawing is also
• don’t use shading covered in Chapter 15, page 264.
• use clear, continuous lines
• use accurate proportions and observation – not a
textbook version.
For a low-power drawing (see Figure 1.7):
• don’t draw individual cells
• draw all tissues completely enclosed by lines
• draw a correct interpretation of the distribution of
tissues
• a representative portion may be drawn (e.g. half a
transverse section).
For a high-power drawing:
• draw only a few representative cells
• draw the cell wall of all plant cells
• don’t draw the nucleus as a solid blob.
Some guidelines for the quality of your labelling:
• label all tissues and relevant structures
Figure 1.7 The right side of this low-power drawing shows
• identify parts correctly examples of good technique, while the left side shows many of
• use a ruler for label lines the pitfalls you should avoid.

Animal and plant cells have
features in common
In animals and plants each cell is surrounded by a very thin
cell surface membrane, which is too thin to be seen with
a light microscope. This is also sometimes referred to as the
plasma membrane.
Many of the cell contents are colourless and transparent so
they need to be stained to be seen. Each cell has a nucleus,
which is a relatively large structure that stains intensely
and is therefore very conspicuous. The deeply staining
material in the nucleus is called chromatin and is a mass
of loosely coiled threads. This material collects together to
form visible separate chromosomes during nuclear division

4
4 1 Cell structure
(see page 86). It contains DNA (deoxyribonucleic acid), a
molecule which contains the instructions that control the
activities of the cell (see Chapter 6). Within the nucleus
an even more deeply staining area is visible, the nucleolus,
which is made of loops of DNA from several chromosomes.
The number of nucleoli is variable, one to five being
common in mammals.
The material between the nucleus and the cell surface
membrane is known as cytoplasm. Cytoplasm is an
aqueous (watery) material, varying from a fluid to a
jelly-like consistency. Many small structures can be seen
within it. These have been likened to small organs and
hence are known as organelles. An organelle can be
defined as a functionally and structurally distinct part
of a cell. Organelles themselves are often surrounded
by membranes so that their activities can be separated
from the surrounding cytoplasm. This is described as
compartmentalisation. Having separate compartments
is essential for a structure as complex as an animal or
plant cell to work efficiently. Since each type of organelle
has its own function, the cell is said to show division of
labour, a sharing of the work between different specialised
organelles.
The most numerous organelles seen with the light
microscope are usually mitochondria (singular:
mitochondrion). Mitochondria are only just visible,
but films of living cells, taken with the aid of a light
microscope, have shown that they can move about, change
shape and divide. They are specialised to carry out aerobic
respiration.
The use of special stains containing silver enabled the
Golgi apparatus to be detected for the first time in 1898
by Camillo Golgi. The Golgi apparatus is part of a complex
internal sorting and distribution system within the cell
(see page 16). It is also sometimes called the Golgi body or
Golgi complex.
Differences between animal
and plant cells
The only structure commonly found in animal cells which
is absent from plant cells is the centriole. Plant cells also
differ from animal cells in possessing cell walls, large
permanent vacuoles and chloroplasts.
Centrioles
Under the light microscope the centriole appears as a small
structure close to the nucleus (see Figure 1.3 on page 2).
The centriole is involved in nuclear division (see page 92).
Cell walls and plasmodesmata
With a light microscope, individual plant cells are more
easily seen than animal cells, because they are usually larger
and, unlike animal cells, surrounded by a cell wall outside
the cell surface membrane. This is relatively rigid because
it contains fibres of cellulose, a polysaccharide which
strengthens the wall. The cell wall gives the cell a definite
shape. It prevents the cell from bursting when water enters
by osmosis, allowing large pressures to develop inside the
cell (see page 77). Cell walls may also be reinforced with
extra cellulose or with a hard material called lignin for
extra strength (see xylem on page 24). Cell walls are freely
permeable, allowing free movement of molecules and ions
through to the cell surface membrane.
Plant cells are linked to neighbouring cells by means of
fine strands of cytoplasm called plasmodesmata (singular:
plasmodesma), which pass through pore-like structures in
the walls of these neighbouring cells. Movement through
the pores is thought to be controlled by the structure of the
pores.
Vacuoles
Although animal cells may possess small vacuoles
such as phagocytic vacuoles (see page 80), which are
temporary structures, mature plant cells often possess a
large, permanent, central vacuole. The plant vacuole is
surrounded by a membrane, the tonoplast, which controls
exchange between the vacuole and the cytoplasm. The fluid
in the vacuole is a solution of mineral salts, sugars, oxygen,
carbon dioxide, pigments, enzymes and other organic
compounds, including some waste products.
Vacuoles help to regulate the osmotic properties of cells
(the flow of water inwards and outwards) as well as having
a wide range of other functions. For example, the pigments
which colour the petals of certain flowers and parts of
some vegetables, such as the red pigment of beetroots, are
sometimes located in vacuoles.
1 Cell structure 5
 Chloroplasts
Some plant cells are able to carry out photosynthesis,
because they contain chloroplasts. Chloroplasts are
relatively large organelles, which are green in colour due to
the presence of chlorophyll. At high magnifications small
‘grains’, or grana (singular: granum), can be seen in the
chloroplasts. During the process of photosynthesis, light is
absorbed by these grana, which actually consist of stacks of
membrane-bound sacs called thylakoids. Starch grains may
also be visible within chloroplasts. Chloroplasts are found
in the green parts of plants, mainly in the leaves.
Points to note
• You can think of a plant cell as being very similar to
an animal cell, but with extra structures.
• Plant cells are often larger than animal cells,
although cell size varies enormously.
• Do not confuse the cell wall with the cell surface
membrane. Cell walls are relatively thick and
physically strong, whereas cell surface membranes
are very thin. Cell walls are freely permeable,
whereas cell surface membranes are partially
permeable. All cells have a cell surface membrane.
• Vacuoles are not confined to plant cells; animal cells
may have small vacuoles, such as phagocytic vacuoles
(see page 80), although these are not usually
permanent structures.
Fraction of a metre
one thousandth   0.001   1/1000   10 -3
one millionth   0.000 001   1/1 000 000   10 -6
one thousand millionth   0.000 000 001   1/1 000 000 000   10
Table 1.1 Units of measurement relevant to cell studies: is the Greek letter mu; 1 micrometre is a thousandth of a millimetre;
1 nanometre is a thousandth of a micrometre.
6
6 1 Cell structure
We return to the differences between animal and plant cells
as seen using the electron microscope on page 18.
Units of measurement in
cell studies
In order to measure objects in the microscopic world, we
need to use very small units of measurement, which are
unfamiliar to most people. According to international
agreement, the International System of Units (SI units)
should be used. In this system the basic unit of length is
the metre (symbol, m). Additional units can be created
in multiples of a thousand times larger or smaller, using
standard prefixes. For example, the prefix kilo means
1000 times. Thus 1 kilometre   1000 metres. The units
of length relevant to cell studies are shown in Table 1.1.
It is difficult to imagine how small these units are,
but, when looking down a microscope and seeing cells
clearly, we should not forget how amazingly small the
cells actually are. The smallest structure visible with the
human eye is about 50–100 m in diameter. Your body
contains about 60 million million cells, varying in size
from about 5 m to 40 m. Try to imagine structures like
mitochondria, which have an average diameter of 1 m.
The smallest cell organelles we deal with in this book,
ribosomes, are only about 25 nm in diameter! You could
line up about 20 000 ribosomes across the full stop at the
end of this sentence.
Unit Symbol
millimetre mm
micrometre m
-9
nanometre nm
Box 1B Measuring cells
Cells and organelles can be measured with a microscope
by means of an eyepiece graticule. This is a transparent a
scale. It usually has 100 divisions (see Figure 1.8a).
The eyepiece graticule is placed in the microscope
eyepiece so that it can be seen at the same time as the
object to be measured, as shown in Figure 1.8b. Figure
0 10 20 30 40 50 60 70 80 90 100
1.8b shows the scale over a human cheek epithelial
cell. The cell lies between 40 and 60 on the scale. We
therefore say it measures 20 eyepiece units in diameter
(the difference between 60 and 40). We will not know
the actual size of the eyepiece units until the eyepiece
graticule scale is calibrated.
To calibrate the eyepiece graticule scale a miniature
transparent ruler called a stage micrometer scale is cheek cells on a slide
placed on the microscope stage and is brought into on the stage of the
focus. This scale may be etched onto a glass slide microscope

or printed on a transparent film. It commonly has b

subdivisions of 0.1 and 0.01 mm. The images of the
two scales can then be superimposed as shown in 0 10 20 30 40 50 60 70 80 90 100
Figure 1.8c.
In the eyepiece graticule shown in the figure,
100 units measure 0.25 mm. Hence, the value of
each eyepiece unit is:
0 25
= 0.0025 mm
100
eyepiece graticule in
Or, converting mm to m: eyepiece graticule the eyepiece of the
scale (arbitrary units) microscope
0 25 × 1000 c
=25 m
100
The diameter of the cell shown superimposed on the
scale in Figure 1.8b measures 20 eyepiece units and so 0 10 20 30 40 50 60 70 80 90 100
its actual diameter is:
20 × 2.5 m 50 m
This diameter is greater than that of many human cells 0 0.1 0.2
because the cell is a flattened epithelial cell.

Figure 1.8 Microscopical measurement. Three fields of view seen

using a high-power ( 40) objective lens. a An eyepiece graticule stage micrometer scale
scale. b Superimposed images of human cheek epithelial cells (marked in 0.0 1mm
and the eyepiece graticule scale. c Superimposed images of the and 0.1 mm divisions)
eyepiece graticule scale and the stage micrometer scale.

1 Cell structure 7
 Electron microscopes
Earlier in this chapter it was stated that by 1900, almost all
the structures shown in Figures 1.3 and 1.5 (pages 2 and 3)
had been discovered. There followed a time of frustration
for microscopists, because they realised that no matter how
much the design of light microscopes improved, there was
a limit to how much could ever be seen using light.
In order to understand the problem, it is necessary to
know something about the nature of light itself and to
understand the difference between magnification and
resolution.
Magnification
Magnification is the number of times larger an image is,
compared with the real size of the object.
observed size of the image
f ation =
magnific
actual size
or
I all measurements to the same units – in this case
M=
A
Worked example 1 – calculating the
magnification of a photograph or object
To calculate M, the magnification of a photograph or
an object, we can use the following method.
Figure 1.9 shows two photographs of a section
through the same plant cells. The magnifications
of the two photographs are the same. Suppose we
want to know the magnification of the plant cell
in Figure 1.9b. If we know its actual (real) length
we can calculate its magnification using the
I
formula M = .
A
The real length of the cell is 80 m.
Step 1
Measure the length in mm of the cell in the photograph
using a ruler. You should find that it is about 60 mm.
Step 2
Convert mm to m. (It is easier if we first convert
micrometres, m.)

where I observed size of the image (that is, what you 1 mm  1000 m
can measure with a ruler) and A actual size (that is, so 60 mm  60 1000 m
the real size – for example, the size of a cell before it is
magnified).  60 000 m
If you know two of these values, you can work out the
third one. For example, if the observed size of the image Step 3
and the magnification are known, you can work out the Use the equation to calculate the magnification.
I
actual size: A = . If you write the formula in a triangle image size, I
M f ation, M =
magnific
as shown below and cover up the value you want to find, it actual size, A
should be obvious how to do the right calculation: 60000 µm
=
80 µm
I
= × 750

The ‘ ’ sign in front of the number 750 means ‘times’.

M A We say that the magnification is ‘times 750’.

Some worked examples are now provided.

8
8 1 Cell structure
 a Worked example 2 – calculating
magnification from a scale bar
Figure 1.10 shows a lymphocyte.

6 µm

Figure 1.10 A lymphocyte.

We can calculate the magnification of the lymphocyte

b by simply using the scale bar. All you need to do
is measure the length of the scale bar and then
substitute this and the length it represents into the
equation.
Step 1
Measure the scale bar. Here, it is 36 mm.
Step 2
Convert mm to m:

36 mm 36 1000 m 36 000 m
Step 3
Use the equation to calculate the magnification:

image size, I
f ation, M =
magnific
actual size, A
36000 µm
=
6 µm
= × 6000
6
Figure 1.9 Photographs of the same plant cells seen a with a
light microscope, b with an electron microscope, both shown at a
magnification of about 750.

1 Cell structure 9

Worked example 3 – calculating the real size
of an object from its magnification
To calculate A, the real or actual size of an object, we
can use the following method.
Figure 1.25 on page 19 shows a plant cell
magnified 5600. One of the chloroplasts is labelled
‘chloroplast’ in the figure. Suppose we want to know
the actual length of this chloroplast.
Step 1
Measure the observed length of the image of the
chloroplast (I ), in mm, using a ruler. The maximum
length is 36 mm.
Step 2
Convert mm to m:
30 mm 30 1000 m 30 000 m
Step 3
Use the equation to calculate the actual length:
image size, I
actual size, A =
magnific
f ation, M
30000 µm
= to colour differences. (Colour is an invention of the brain!)
5600
= 5 4 µm (to one decimal place)
SAQ 1.2
a Calculate the magnification of the drawing of the animal
cell in Figure 1.3 on page 2.
b Calculate the actual (real) length of the bottom
chloroplast in Figure 1.27 on page 19.
Resolution
Look again at Figure 1.9 (page 9). Figure 1.9a is a light
micrograph (a photograph taken with a light microscope,
also known as a photomicrograph). Figure 1.9b is an
electron micrograph of the same cells taken at the same
magnification (an electron micrograph is a picture taken
with an electron microscope). You can see that Figure 1.9b,
the electron micrograph, is much clearer. This is because it
has greater resolution. Resolution is defined as the ability
010
1 1 Cell structure
to distinguish between two separate points. If the two
points cannot be resolved, they will be seen as one point.
In practice, resolution is the amount of detail that can be
seen – the greater the resolution, the greater the detail.
The maximum resolution of a light microscope is 200 nm.
This means that if two points or objects are closer together
than 200 nm they cannot be distinguished as separate.
It is possible to take a photograph such as Figure 1.9a
and to magnify (enlarge) it, but we see no more detail; in
other words, we do not improve resolution, even though
we often enlarge photographs because they are easier to see
when larger. With a microscope, magnification up to the
limit of resolution can reveal further detail, but any further
magnification increases blurring as well as the size of the
image.
The electromagnetic spectrum
How is resolution linked with the nature of light? One
of the properties of light is that it travels in waves. The
length of the waves of visible light varies, ranging from
about 400 nm (violet light) to about 700 nm (red light).
The human eye can distinguish between these different
wavelengths, and in the brain the differences are converted
The whole range of different wavelengths is called the
electromagnetic spectrum. Visible light is only one part of
this spectrum. Figure 1.11 shows some of the parts of the
electromagnetic spectrum. The longer the waves, the lower
their frequency (all the waves travel at the same speed, so
imagine them passing a post: shorter waves pass at higher
frequency). In theory, there is no limit to how short or how
long the waves can be. Wavelength changes with energy: the
greater the energy, the shorter the wavelength (rather like
squashing a spring).
Now look at Figure 1.12, which shows a mitochondrion,
some very small cell organelles called ribosomes (see
page 13) and light of 400 nm wavelength, the shortest
visible wavelength. The mitochondrion is large enough
to interfere with the light waves. However, the ribosomes
are far too small to have any effect on the light waves. The
general rule is that the limit of resolution is about one half
the wavelength of the radiation used to view the specimen.
In other words, if an object is any smaller than half the
wavelength of the radiation used to view it, it cannot be
seen separately from nearby objects. This means that the
 X-rays infrared microwaves
v
gamma rays i
s radio and TV waves
uv
i
b
l
0.1 nm 10 nm e 1000 nm 105 nm 107 nm 109 nm 1011 nm 1013 nm

visible light

400 nm 500 nm 600 nm 700 nm

violet blue green yellow orange red

Figure 1.11 Diagram of the electromagnetic spectrum (the waves are not drawn to scale). The numbers indicate the wavelengths of the different
types of electromagnetic radiation. Visible light is a form of electromagnetic radiation.

best resolution that can be obtained using a microscope
that uses visible light (a light microscope) is 200 nm,
since the shortest wavelength of visible light is 400 nm
(violet light). In practice, this corresponds to a maximum
useful magnification of about 1500 times. Ribosomes are
approximately 25 nm in diameter and can therefore never
be seen using light.
If an object is transparent, it will allow light waves to pass
through it and therefore will still not be visible. This is why
many biological structures have to be stained before they
can be seen.
wavelength stained mitochondrion
400 nm of diameter 1000 nm
interferes with light waves
stained ribosomes of diameter 25 nm
do not interfere with light waves
Figure 1.12 A mitochondrion and some ribosomes in the path of light
waves of 400 nm length.
The electron microscope
Biologists, faced with the problem that they would never
see anything smaller than 200 nm using a light microscope,
realised that the only solution would be to use radiation of
a shorter wavelength than light. If you study Figure 1.11,
you will see that ultraviolet light, or better still X-rays,
look like possible candidates. Both ultraviolet and X-ray
microscopes have been built, the latter with little success
partly because of the difficulty of focusing X-rays. A much
better solution is to use electrons. Electrons are negatively
charged particles which orbit the nucleus of an atom.
When a metal becomes very hot, some of its electrons
gain so much energy that they escape from their orbits,
like a rocket escaping from Earth’s gravity. Free electrons
behave like electromagnetic radiation. They have a very
short wavelength: the greater the energy, the shorter the
wavelength. Electrons are a very suitable form of radiation
for microscopy for two major reasons. Firstly, their
wavelength is extremely short (at least as short as that of
X-rays). Secondly, because they are negatively charged, they
can be focused easily using electromagnets (a magnet can be
made to alter the path of the beam, the equivalent of a glass
lens bending light).
Using an electron microscope, a resolution of 0.5 nm
can be obtained, 400 times better than when using a light
microscope.
Transmission and scanning electron microscopes E
Two types of electron microscope are now in common
use. The transmission electron microscope, or TEM for

1 Cell structure 11
E E
short, was the type originally developed. Here the beam
electron gun and anode, which
of electrons is passed through the specimen before being produce a beam of electrons
viewed. Only those electrons that are transmitted (pass
through the specimen) are seen. This allows us to see thin electron beam
sections of specimens, and thus to see inside cells. In the
scanning electron microscope (SEM), on the other hand, vacuum
the electron beam is used to scan the surfaces of structures,
pathway of electrons
and only the reflected beam is observed.
An example of a scanning electron micrograph is shown
in Figure 1.13. The advantage of this microscope is that condenser electromagnetic
lens, which directs the electron
surface structures can be seen. Also, great depth of field
beam onto the specimen
is obtained so that much of the specimen is in focus at
the same time and a three-dimensional appearance is
obtained. Such a picture would be impossible to obtain
specimen, which is
with a light microscope, even using the same magnification placed on a grid
and resolution, because you would have to keep focusing
up and down with the objective lens to see different parts
of the specimen. The disadvantage of the SEM is that it objective electromagnetic
cannot achieve the same resolution as a TEM. Resolution is lens, which produces an
image
between 3 nm and 20 nm.
Viewing specimens with the electron microscope
Figure 1.14 shows how an electron microscope works projector electromagnetic
and Figure 1.15 shows one in use. lenses, which focus the
magnified image onto
the screen

screen or photographic
plate, which shows the
image of the specimen

Figure 1.14 How an electron microscope works.

It is not possible to see an electron beam, so to make the

Figure 1.13 False-colour SEM of the head of a cat flea ( 100).
image visible the electron beam has to be projected onto a
fluorescent screen. The areas hit by electrons shine brightly,
giving overall a ‘black and white’ picture. The stains used
to improve the contrast of biological specimens for electron
microscopy contain heavy metal atoms, which stop the
passage of electrons. The resulting picture is like an X-ray
photograph, with the more densely stained parts of the
specimen appearing blacker. ‘False-colour’ images can be
created by colouring the standard black and white image
using a computer.
To add to the difficulties of electron microscopy, the
electron beam, and therefore the specimen and the
fluorescent screen, must be in a vacuum. If electrons

212
1 1 Cell structure

E organelles
Figure 1.15 A TEM in use.
collided with air molecules, they would scatter, making it
impossible to achieve a sharp picture. Also, water boils at
room temperature in a vacuum, so all specimens must be
dehydrated before being placed in the microscope. This
means that only dead material can be examined. Great
efforts are therefore made to try to preserve material in a
life-like state when preparing it for the microscope.
SAQ 1.3
Explain why ribosomes are not visible using a light
microscope.
Ultrastructure of an animal cell
The ‘fine’, or detailed, structure of a cell as revealed by the
electron microscope is called its ultrastructure. Figure 1.16
shows the appearance of typical animal cells as seen with
an electron microscope, and Figure 1.17 on page 15 is a
diagram based on many other such micrographs.
SAQ 1.4
Compare Figure 1.17 on page 15 with Figure 1.3 on
page 2. Name the structures which can be seen with the
electron microscope but not with the light microscope.
Structures and functions of
Compartmentalisation and division of labour within the
cell are even more obvious with an electron microscope
than with a light microscope.
We will now consider the structures and functions of
some of the cell components in more detail.
Nucleus
The nucleus (Figure 1.18 on page 15) is the largest
cell organelle (see also page 5). It is surrounded by two
membranes known as the nuclear envelope. The outer
membrane of the nuclear envelope is continuous with
the endoplasmic reticulum (Figure 1.17 on page 15). The
nuclear envelope has many small pores called nuclear
pores. These allow and control exchange between the
nucleus and the cytoplasm. Examples of substances leaving
the nucleus through the pores are mRNA and ribosomes for
protein synthesis. Examples of substances entering through
the nuclear pores are proteins to help make ribosomes,
nucleotides, ATP (aderosine triphosphate) and some
hormones such as thyroid hormone T3.
Within the nucleus, the chromosomes are in a loosely
coiled state known as chromatin (except during nuclear
division, see Chapter 5). Chromosomes contain DNA,
which is organised into functional units called genes. Genes
control the activities of the cell and inheritance; thus the
nucleus controls the cell’s activities. When a cell is about
to divide, the nucleus divides first so that each new cell
will have its own nucleus (Chapters 5 and 19). Also within
the nucleus, the nucleolus makes ribosomes, using the
information in its own DNA.
Endoplasmic reticulum and ribosomes
When cells were first seen with the electron microscope,
biologists were amazed to see so much detailed structure.
The existence of much of this had not been suspected. This
was particularly true of an extensive system of membranes
running through the cytoplasm, which became known
as the endoplasmic reticulum (ER) (Figure 1.19 on
page 15 – see also Figures 1.18 on page 15 and 1.22 on
page 17). The ER is continuous with the outer membrane
of the nuclear envelope (Figure 1.17).
There are two types of ER: rough ER and smooth ER.
Rough ER is so called because it is covered with many tiny
1 Cell structure 13
 lysosome
Golgi apparatus
cell surface
membrane

mitochondria

nucleolus

endoplasmic
reticulum
nucleus

glycogen granules

microvillus chromatin

nuclear envelope

ribosomes

Figure 1.16 Representative animal cells as seen with a TEM. The cells are liver cells from a rat ( 9600). The nucleus is clearly visible in one of the cells.

414
1 1 Cell structure
 two centrioles close to the
nucleus and at right angles microvilli
to each other

mitochondrion Golgi vesicle

lysosome Golgi apparatus

ribosomes

rough
endoplasmic cell surface
reticulum membrane

nucleolus
chromatin smooth endoplasmic cytoplasm
nucleus nuclear pore reticulum

nuclear envelope
(two membranes)

Figure 1.17 Ultrastructure of a typical animal cell as seen with an electron microscope. In reality, the ER is more extensive than shown, and free
ribosomes may be more extensive. Glycogen granules are sometimes present in the cytoplasm.

Figure 1.18 TEM of the nucleus of a cell from the pancreas of a bat
( 7500). The circular nucleus is surrounded by a double-layered
nuclear envelope containing nuclear pores. The nucleolus is more Figure 1.19 TEM of rough ER covered with ribosomes (black dots)
darkly stained. Rough ER is visible in the surrounding cytoplasm. ( 17 000). Some free ribosomes can also be seen in the cytoplasm.

1 Cell structure 15
 organelles called ribosomes. These are just visible as black
dots in Figures 1.18 and 1.19 on page 15. At very high
magnifications they can be seen to consist of two subunits:
a large and a small subunit. Ribosomes are the sites of
protein synthesis (see pages 111–112). They can be found
free in the cytoplasm as well as on the rough ER. They are
very small, only about 25 nm in diameter. They are made
of RNA (ribonucleic acid) and protein. The rough ER
forms an extensive system of flattened sacs spreading in
sheets throughout the cell. Proteins made by the ribosomes
on the rough ER enter the sacs and move through them.
The proteins are often processed in some way on their
journey. Small sacs called vesicles can break off from the ER
and these can join together to form the Golgi apparatus.
Proteins can be exported from the cell via the Golgi
apparatus (see page 80).
Smooth ER, so called because it lacks ribosomes, has
a completely different function. It makes lipids and
steroids, such as cholesterol and the reproductive hormones
oestrogen and testosterone.
Golgi apparatus (Golgi body or Golgi complex)
The Golgi apparatus is a stack of flattened sacs (Figure
1.20). This stack of sacs is sometimes referred to as the
Figure 1.20 TEM of a Golgi apparatus. A central stack of saucer-shaped
sacs can be seen budding off small Golgi vesicles (green). These may
form secretory vesicles whose contents can be released at the cell
surface by exocytosis (see page 80).
616
1 1 Cell structure
Golgi body. More than one may be present in a cell. The
stack is constantly being formed at one end from vesicles
which bud off from the ER, and broken down again at the
other end to form Golgi vesicles. The stack of sacs with the
associated vesicles is referred to as the Golgi apparatus or
Golgi complex.
The Golgi apparatus collects, processes and sorts
molecules (particularly proteins from the rough ER), ready
for transport in Golgi vesicles either to other parts of the
cell or out of the cell (secretion). Two examples of protein
processing in the Golgi apparatus are the addition of sugars
to proteins to make molecules known as glycoproteins,
and the removal of the first amino acid, methionine, from
newly formed proteins to make a functioning protein. In
plants, enzymes in the Golgi apparatus convert sugars into
cell wall components. Golgi vesicles are also used to make
lysosomes.
Lysosomes
Lysosomes (Figure 1.21) are spherical sacs, surrounded
by a single membrane and having no internal structure.
They are commonly 0.1– 0.5 m in diameter. They contain
digestive (hydrolytic) enzymes which must be kept separate
Figure 1.21 Lysosomes (orange) in a mouse kidney cell ( 55 000). They
contain cell structures in the process of digestion and vesicles (green).
Cytoplasm is coloured blue here.
from the rest of the cell to prevent damage. Lysosomes are
responsible for the breakdown (digestion) of unwanted
structures such as old organelles or even whole cells, as
in mammary glands after lactation (breast feeding). In
white blood cells, lysosomes are used to digest bacteria
(see endocytosis, page 80). Enzymes are sometimes
released outside the cell – for example, in the replacement
of cartilage with bone during development. The heads
of sperm contain a special lysosome, the acrosome, for
digesting a path to the ovum (egg).
Mitochondria
Mitochondria (singular: mitochondrion) are usually about
1 m in diameter and can be various shapes, often sausage-
shaped as in Figure 1.22. They are surrounded by two
membranes (an envelope). The inner of these is folded
to form finger-like cristae which project into the interior
solution, or matrix.
The main function of mitochondria is to carry out aerobic
respiration. As a result of respiration, they make ATP, the
universal energy carrier in cells (see Chapter 16). They are
also involved in the synthesis of lipids (page 37).
Figure 1.22 Mitochondrion (orange) with its double membrane
(envelope); the inner membrane is folded to form cristae ( 20 000).
Mitochondria are the sites of aerobic cell respiration. Note also the
rough ER.
In the 1960s it was discovered that mitochondria and
chloroplasts contain ribosomes which are slightly smaller
than those in the cytoplasm and are the same size as those
found in bacteria. The size of ribosomes is measured in ‘S
units’, which are a measure of how fast they sediment in a
centrifuge. Cytoplasmic ribosomes are 80S, while those of
bacteria, mitochondria and chloroplasts are 70S. It was also
discovered in the 1960s that mitochondria and chloroplasts
contain small, circular DNA molecules, also like those
found in bacteria. Not surprisingly, it was later proved
that mitochondria and chloroplasts are, in effect, ancient
bacteria which now live inside the larger cells typical of
animals and plants (see prokaryotic and eukaryotic cells,
page 18). This is known as the endosymbiont theory.
‘Endo’ means ‘inside’ and a ‘symbiont’ is an organism which
lives in a mutually beneficial relationship with another
organism. The DNA and ribosomes of mitochondria and
chloroplasts are still active and responsible for the coding
and synthesis of certain vital proteins, but mitochondria
and chloroplasts can no longer live independently.
Mitochondrial ribosomes are just visible as tiny dark dots
in the mitochondrial matrix in Figure 1.22.
Cell surface membrane
The cell surface membrane is extremely thin (about 7 nm).
However, at very high magnifications, at least 100 000, it
can be seen to have three layers, described as a trilaminar
appearance. This consists of two dark lines (heavily stained)
either side of a narrow, pale interior (Figure 1.23). The
membrane is partially permeable and controls exchange
between the cell and its environment. Membrane structure
is discussed further in Chapter 4.
Figure 1.23 Cell surface membrane ( 250 000). At this magnification
the membrane appears as two dark lines at the edge of the cell.
Microvilli
Microvilli (singular: microvillus) are finger-like extensions
of the cell surface membrane, typical of certain epithelial
cells (cells covering surfaces of structures). They greatly
1 Cell structure 17
 increase the surface area of the cell surface membrane (see
Figure 1.17 on page 15). This is useful, for example, for
absorption in the gut and for reabsorption in the proximal
convoluted tubules of the kidney (see page 307).
Centrioles
The extra resolution of the electron microscope reveals
that just outside the nucleus, there are really two centrioles
(see Figure 1.24), not one as it appears under the light
microscope (compare with Figure 1.3 on page 2). They lie
close together at right-angles to each other. A centriole is
a hollow cylinder about 0.4 m long, formed from a ring
of short microtubules, tiny tubes made of a protein called
tubulin. These microtubules are used as a starting point for
growing the spindle microtubules for nuclear division (see
page 92). Centrioles are not found in plant cells.
Ultrastructure of a plant cell
All the structures except centrioles and microvilli so far
described in animal cells are also found in plant cells.
The appearance of a plant cell as seen with the electron
microscope is shown in Figure 1.25 while Figure 1.26 is a
diagram based on many such micrographs. The relatively
thick cell wall and the large central vacuole are obvious,
as are the chloroplasts, two of which are shown in detail
in Figure 1.27. These structures and their functions have
been described on pages 5 and 6. The electron microscope
Figure 1.24 Centrioles in transverse and longitudinal section (TS and
LS) ( 86 000). The one on the left is seen in TS and clearly shows the
nine triplets of microtubules which make up the structure.
818
1 1 Cell structure
reveals that chloroplasts contain 70S ribosomes and small,
circular DNA molecules, as do mitochondria and bacteria.
This has already been discussed with mitochondria above
(page 17).
SAQ 1.5
Compare Figure 1.26 with Figure 1.5 on page 3. Name
the structures which can be seen with the electron
microscope but not with the light microscope.
Two fundamentally different
types of cell
At one time it was common practice to try to classify
all living organisms as either animals or plants. With
advances in our knowledge of living things, it has
become obvious that the living world is not that simple.
Fungi and bacteria, for example, are very different from
animals and plants, and from each other. Eventually it
was discovered that there are two fundamentally different
types of cell. The most obvious difference between
these types is that one possesses a nucleus and the other
does not.
Organisms that lack nuclei are called prokaryotes (‘pro’
means before; ‘karyon’ means nucleus). All prokaryotes are
now referred to as bacteria. They are, on average, about
1000 to 10 000 times smaller in volume than cells with
nuclei, and are much simpler in structure – for example,
their DNA lies free in the cytoplasm.
Organisms whose cells possess nuclei are called
eukaryotes (‘eu’ means true). Their DNA lies inside a
nucleus. Eukaryotes include animals, plants, fungi and a
group containing most of the unicellular eukaryotes known
as protoctists. Most biologists believe that eukaryotes
evolved from prokaryotes, one-and-a-half thousand million
years after prokaryotes first appeared on Earth. We mainly
study animals and plants in this book, but all eukaryotic
cells have certain features in common.
A generalised prokaryotic cell is shown in Figure 1.28. A
comparison of prokaryotic and eukaryotic cells is given in
Table 1.2 on page 21.