GAS EXCHANGE

Gas Exchange in Organisms

• Cellular respiration is a process occurring in all living cells that releases energy in
the form of ATP
o This energy is released when substrate molecules such as glucose is oxidised
o Organisms use this energy to perform important life functions such as
nutrition and excretion
• Aerobic respiration requires oxygen to occur and it produces carbon dioxide as a
waste product
o Living organisms acquire this oxygen from their environment and release
carbon dioxide back into their surroundings
• The process by which these gases are exchanged between living organisms and their
environment is called gas exchange
o This includes oxygen uptake and the release of carbon dioxide by organisms
o In plants, carbon dioxide will be absorbed and oxygen released during the day
as a result of photosynthesis
• Gas exchange takes place by the process of diffusion, the rate of which is determined
by the following factors:
o Size of the respiratory surface - the bigger the surface, the higher the rate of
diffusion
o Concentration gradient
o Diffusion distance - the shorter the distance, the higher the rate of diffusion
• Small, unicellular organisms such as amoeba have a large surface area compared to
the volume of cytoplasm and a short diffusion distance
o This means that the rate of diffusion is sufficient to supply the organism with
enough oxygen to function

Single Celled Organism Diffusion Diagram

Small, unicellular organisms have a large surface area to volume ratio and a short
diffusion distance to allow for effective gas exchange to occur
Challenges of gas exchange in organisms

• As an organism increases in size, the challenges of gas exchange become greater

• This is because an increase in size will result in a:
o Smaller surface area to volume ratio
o Greater diffusion distance
• Large, multicellular organisms therefore cannot rely on diffusion alone to supply
every cell with oxygen
o Another challenge is that the external surface of these organisms are
designed to provide protection to the tissue underneath and is therefore not
suitable as a respiratory surface
• The cells of large, active organisms will require more oxygen than smaller, less
active organisms in order to meet their metabolic demands
o These organisms will require specialised organs for gas exchange

Gas Exchange Surfaces: Properties

• To maximise the rate of diffusion of oxygen and carbon dioxide, gas exchange
surfaces require certain properties which include:
o Permeability in order for gases to move across the surface
o Thin tissue layer to create a short diffusion distance for oxygen and carbon
dioxide
o Presence of moisture so that gases can dissolve
§ This will facilitate the diffusion of gases across a gas exchange surface
o Large surface area so that many gas molecules can diffuse across at the same
time

Maintaining a Concentration Gradient

• A steep concentration gradient will ensure a high diffusion rate across a gas
exchange surface
o In organisms, this will allow the diffusion of oxygen into the body and the
diffusion of carbon dioxide out of the body
• These concentration gradients are maintained in the following ways:
o A dense network of blood vessels to provide a large surface area for the
diffusion of gases
§ Blood provides a good transport medium for both oxygen and carbon
dioxide
o A continuous blood flow in the blood vessels to ensure that oxygen is
constantly transported away from the gas exchange surface and carbon dioxide
towards them
§ This ensures that oxygen will always diffuse into the blood and carbon
dioxide out of the blood in the lungs
o Ventilation with air in lungs and water in gills to bring oxygen close to the
gas exchange surface and to remove carbon dioxide
Alveolus Diagram

The alveolus is the gas exchange surface in humans where a concentration gradient for
oxygen and carbon dioxide is maintained

Mammalian Lungs: Adaptations

• Air moves in through the nose and mouth before it is carried to the lungs through
the trachea
• The trachea is a tube supported by rings of cartilage which help to support its shape
and ensure it stays open while allowing it to move and flex with the body
• The trachea divides to form the two bronchi (singular bronchus) with walls also
strengthened with cartilage and a layer of smooth muscle that can contract or relax to
change the diameter of the airways. Both trachea and bronchi are lined with ciliated
epithelium to remove particles trapped in mucus that enter the airways
o One bronchus leads to each lung
• Bronchioles branch off the two bronchi to form a network of narrow tubes
o The walls of the bronchioles are lined with a layer of smooth muscle to alter
the diameter of the bronchiole tubes
o This helps to regulate the flow of air into the lungs by dilating when more air
is needed and constricting when e.g. an allergen is present
 • Groups of alveoli are found at the end of the bronchioles
• Each alveolus is surrounded by an extensive network of capillaries to provide a good
blood supply for maximum gas exchange

Human Gas Exchange System Diagram

The main structures of the human gas exchange system

Adaptations of mammalian lungs for gas exchange

• Each mammalian lung is comprised of many, small alveoli

o These provide a large surface area for gas exchange
• Alveoli are grouped around the ends of bronchioles, which spreads out to form
a branched network across each lung
o This ensures an even distribution of alveoli throughout the lungs
• The clusters of alveoli are surrounded by an extensive capillary bed
o This provides an increased surface area for the diffusion of oxygen and
carbon dioxide between the alveoli and blood
o Deoxygenated blood enters the capillary beds from a branch of the pulmonary
artery while oxygenated blood leaves the capillary beds via a branch of the
pulmonary vein
§ This maintains the concentration gradient of oxygen and carbon
dioxide between the alveoli and blood
 • Cells of the alveolar wall secrete a substance called surfactant which lowers the
surface tension in the alveoli
o This prevents the alveoli from collapsing and sticking together during
expiration

Human Alveoli Diagram

Many, small alveoli and an extensive capillary network are examples of how the
mammalian lung is adapted for gas exchange
Ventilation: Mechanism
• Ventilation is essential for the effective exchange of gases in the lungs
o It replaces older air in the lungs with fresh air from the external environment
o This helps to maintain the concentration gradient of oxygen and carbon
dioxide between the alveoli and blood
• Ventilation involves inspiration (breathing in) and expiration (breathing out)

Inspiration

• The breathing-in, or inspiration, process causes the volume of the chest to

increase and the air pressure to decrease until it is lower than the atmospheric
pressure
o When gas is in a large volume container that allows the gas particles to spread
out, the pressure exerted by the gas on the walls of the container is low
• As a result, air moves down the pressure gradient and rushes into the lungs
o A gas will always move down a pressure gradient from an area of high
pressure to an area of low pressure
• The inspiration process
o The diaphragm contracts and flattens, increasing chest volume
o In addition to the flattening of the diaphragm the external intercostal
muscles contract, causing the ribcage to move upwards and outwards; this
also increases chest volume
 The process of inspiration

Expiration

• Breathing out, or expiration, occurs mostly due to the recoil of the lungs after they
have been stretched by the inspiration process, and is therefore a mainly passive
process
• Volume of the chest decreases and pressure increases, causing air to be forced out
down its pressure gradient
o When gas is in a low volume container it is compressed, causing the gas
particles to exert more pressure on the walls of the container
• The passive expiration process
o External intercostal muscles relax, allowing the ribcage to move down and in
o Diaphragm relaxes and becomes dome-shaped
 o The recoil of elastic fibres in the alveoli walls reduces the volume of the
lungs
• The expiration process can be active when there is a need to expel excess air from the
lungs e.g. when blowing out a candle
• The active expiration process
o Internal intercostal muscles contract to pull the ribs down and in
o Abdominal muscles contract to push organs upwards against the diaphragm,
decreasing the volume of the chest cavity
o This causes forced exhalation

Haemoglobin & Oxygen

Foetal & Adult Haemoglobin
 • Haemoglobin is the molecule responsible for binding oxygen in our blood
o They are globular proteins found in abundance in red blood cells
o Each haemoglobin molecule consists of four polypeptide subunits
o At the centre of each subunit is an iron-containing haem group with which
oxygen combines
§ Each haem group can bind to one oxygen molecule
§ That means that each molecule of haemoglobin can
transport four oxygen molecules
• Oxygen is one of the gases found in air and each of these gases exerts a pressure
o The pressure of each gas in a mixture of gases is called its partial pressure
o The symbol for partial pressure is p, therefore the partial pressure of oxygen
can be denoted as pO2
• Due to the shape of the haemoglobin molecule it is difficult for the first oxygen
molecule to bind to its haem group
• However, after the first oxygen molecule binds, the haemoglobin protein changes
shape, or conformation, making it easier for the next oxygen molecules to bind
o This is known as cooperative binding
• The ease with which haemoglobin binds and dissociates with oxygen can be described
as its affinity for oxygen
o In areas where there are high partial pressures of oxygen (such as the alveoli
of the lungs), the affinity of haemoglobin for oxygen is high
§ This means haemoglobin and oxygen will bind easily
o In areas where there are low partial pressures of oxygen (such as respiring
muscle cells), the affinity of haemoglobin for oxygen is low
§ This means haemoglobin and oxygen will dissociate easily from each
other
o This ensures that haemoglobin can easily bind to oxygen in the lung
capillaries to transport it to and then release it near respiring cells that require
oxygen

Foetal haemoglobin

• The haemoglobin of a developing foetus has a higher affinity for oxygen than adult
haemoglobin
• This is vital as it allows a foetus to obtain oxygen from its mother's blood at the
placenta
o Foetal haemoglobin can bind to oxygen at low pO2
o At this low pO2 the mother's haemoglobin is dissociating with oxygen
• We can represent the percentage saturation of haemoglobin at different partial
pressures of oxygen as a graph
o This is called the oxygen dissociation curve
• On a dissociation curve, the curve for foetal haemoglobin shifts to the left of that for
adult haemoglobin
o This means that at any given partial pressure of oxygen, foetal
haemoglobin has a higher percentage saturation than adult haemoglobin
• After birth, a baby begins to produce adult haemoglobin which gradually
replaces foetal haemoglobin
o This is important for the easy release of oxygen in the respiring tissues of a
more metabolically active individual
 Foetal haemoglobin has a higher affinity for oxygen; its oxygen dissociation curve
therefore lies further to the left than the curve of adult haemoglobin

• Haemoglobin has the ability to change shape, or conformation, once oxygen binds to
it due to cooperative binding
o Proteins like this are known as allosteric proteins as they can exist in
multiple conformations
• Carbon dioxide is an allosteric inhibitor of haemoglobin
o This means that when it binds to haemoglobin, it is more difficult for oxygen
to bind to haemoglobin as the protein cannot change its conformation
o This lowers the affinity of haemoglobin for oxygen
• Carbon dioxide has less of an allosteric effect on foetal haemoglobin
o This enables foetal haemoglobin to have a higher affinity for oxygen even if
carbon dioxide is bound to it

The Bohr Shift

• Changes in the oxygen dissociation curve as a result of carbon dioxide levels are
known as the Bohr effect, or Bohr shift
• When the partial pressure of carbon dioxide in the blood is high, haemoglobin’s
affinity for oxygen is reduced
o This is the case in respiring tissues, where cells are producing carbon dioxide
as a waste product of respiration
o This occurs because CO2 lowers the pH of the blood
§ CO2 combines with water to form carbonic acid
 § Carbonic acid dissociates into hydrogen carbonate ions and hydrogen
ions
§ Hydrogen ions bind to haemoglobin, causing the release of oxygen
• This is a helpful change because it means that haemoglobin gives up its oxygen
more readily in the respiring tissues where it is needed
• On a graph showing the dissociation curve, the curve shifts to the right when
CO2 levels increase
o This means that at any given partial pressure of oxygen, the percentage
saturation of haemoglobin is lower at higher levels of CO2

The dissociation curve shifts to the right as a result of the Bohr effect. This means that any
given partial pressure of oxygen, the percentage saturation of haemoglobin is lower at
higher CO2 levels.

The Oxygen Dissociation Curve

• The oxygen dissociation curve shows the rate at which oxygen associates, and also
dissociates, with haemoglobin at different partial pressures of oxygen (pO2)
o Partial pressure of oxygen refers to the pressure exerted by oxygen within a
mixture of gases; it is a measure of oxygen concentration
o Haemoglobin is referred to as being saturated when all of its oxygen
binding sites are taken up with oxygen; so when it contains four oxygen
molecules
• The ease with which haemoglobin binds and dissociates with oxygen can be described
as its affinity for oxygen
o When haemoglobin has a high affinity it binds easily and dissociates slowly
o When haemoglobin has a low affinity for oxygen it binds
slowly and dissociates easily
• In other liquids, such as water, we would expect oxygen to becomes associated with
water, or to dissolve, at a constant rate, providing a straight line on a graph, but
with haemoglobin oxygen binds at different rates as the pO2 changes; hence the
resulting curve
o It can be said that haemoglobin's affinity for oxygen changes at different
partial pressures of oxygen
 The oxygen dissociation curve

Interpreting the curve

• When the curve is read from left to right, it provides information about the rate at
which haemoglobin binds to oxygen at different partial pressures of oxygen
o At low pO2 (the bottom left corner of the graph) oxygen binds slowly to
haemoglobin; this means that haemoglobin cannot pick up oxygen and
become saturated as blood passes through the body's oxygen-depleted tissues
§ Haemoglobin has a low affinity for oxygen at low pO2, so saturation
percentage is low
 o At medium pO2 (in the central region of the graph) oxygen binds more easily
to haemoglobin and saturation increases quickly; at this point on the graph
a small increase in pO2 causes a large increase in haemoglobin saturation
o At high pO2 (in the top right corner of the graph) oxygen binds easily to
haemoglobin; this means that haemoglobin can pick up oxygen and become
saturated as blood passes through the lungs
§ Haemoglobin has a high affinity for oxygen at high pO2, so saturation
percentage is high
§ Note that at this point on the graph increasing the pO2 by a large
amount only has a small effect on the percentage saturation of
haemoglobin; this is because most oxygen binding sites on
haemoglobin are already occupied
• When read from right to left, the curve provides information about the rate at which
haemoglobin dissociates with oxygen at different partial pressures of oxygen
o In the lungs, where pO2 is high, there is very little dissociation of oxygen
from haemoglobin
o At medium pO2, oxygen dissociates readily from haemoglobin, as shown by
the steep region of the curve; this region corresponds with the partial
pressures of oxygen present in the respiring tissues of the body, so ready
release of oxygen is important for cellular respiration
§ At this point on the graph a small decrease in pO2 causes a large
decrease in percentage saturation of haemoglobin, leading to easy
release of plenty of oxygen to the cells
o At low pO2 dissociation slows again; there are few oxygen molecules left on
the binding sites, and the release of the final oxygen molecule becomes more
difficult, in a similar way to the slow binding of the first oxygen molecule

Explaining the curve

• The curved shape of the oxygen dissociation curve for haemoglobin can be explained
as follows
o Due to the shape of the haemoglobin molecule it is difficult for the first
oxygen molecule to bind to haemoglobin
§ This means that binding of the first oxygen occurs slowly, explaining
the relatively shallow curve at the bottom left corner of the graph
o After the first oxygen molecule binds to haemoglobin, the haemoglobin
protein changes shape, or conformation, making it easier for the next
oxygen molecules to bind due to cooperative binding
§ This speeds up binding of the remaining oxygen molecules and
explains the steeper part of the curve in the middle of the graph
o As the haemoglobin molecule approaches saturation it takes longer for the
fourth oxygen molecule to bind
§ This is due to the shortage of remaining binding sites, explaining
the levelling off of the curve in the top right corner of the graph

Measuring Lung Volumes

• It is possible to investigate the effect of exercise on ventilation using an apparatus
called a spirometer
 o It contains a chamber filled with water which is covered by a hinged plastic lid
o The person partaking in the experiment breathes through a mouthpiece which
is connected to the spirometer chamber
o The plastic lid moves up and down as breathing occurs
• The spirometer chamber could be filled with either air or oxygen
o When filled with air, it can be used to determine lung capacity in different
conditions
o When filled with oxygen and soda lime (for absorbing carbon dioxide), it can
measure oxygen consumption in different conditions
• Spirometer traces are created by:
o Drawing a line on a revolving drum as the lid moves
o A computer which draws a graph of the results
• Several measurements can be made using spirometer traces such as:
o Ventilation rate
o Tidal volume
o Reserve volumes during inspiration and expiration
o Vital capacity

A classic spirometer can be used to investigate ventilation

 Using a spirometer to monitor ventilation can also be carried out with an electric
spirometer

Analysis of spirometer trace

• The effect of exercise on ventilation can be seen in the spirometer trace below
Tidal volume

• The tidal volume is the volume of air inhaled and exhaled during normal breathing
o Exercise will lead to an increase in the tidal volume as more air is moved in
and out of the lungs
o We do have the potential to take extra deep breaths
§ The maximum volume of air that can enter the lungs during inspiration
is known as the maximum inspiratory level
§ Similarly, the maximum volume of air that can be exhaled during
expiration is known as the maximum expiratory level

Inspiratory and expiratory reserve volumes

• The reserve volumes of the lungs refer to the extra volume of air that can be inhaled
or exhaled when taking an extra deep breath and are determined as follows:
o The difference between the maximum inspiratory level and tidal volume is
called the inspiratory reserve volume
 o The difference between the maximum expiratory level and tidal volume is
called the expiratory reserve volume

Vital capacity

• The vital capacity (VC) refers to the total amount of air exhaled after taking a deep
breath
o This can be calculated by adding the tidal volume (TV), inspiratory reserve
volume (IRV) and expiratory reserve volume (ERV) together

VC = TV + IRV + ERV

Ventilation rate

• The ventilation rate can be determined by counting the number of inhalations or

exhalations per minute
o Exercise will cause an increase in the ventilation rate as you will be taking
more breaths per minute

Gas Exchange in Plants

Leaf Adaptations for Gas Exchange
• Gas exchange in plants occur through the leaf
• The leaf contains the following tissues:
o Epidermal tissue forming the outer boundary of the leaf
o Mesophyll tissue that make up the bulk of internal structure of the leaf
o Vascular tissue which transports substances between the leaf and the rest of
the plant

Epidermis

• This is formed by a single layer of tightly packed cells

o The leaf has an upper and lower epidermis which protects the inner parts of
the leaf
• The lower epidermis contains tiny pores called stomata (singular stoma)
o Each stoma is surrounded by two guard cells which controls the opening and
closure of the pore
§ When water moves into the guard cells they become turgid and
change shape which opens the stomata
§ They become flaccid when water is lost and this causes the stomata to
close
o Stomata are the structures through which gas exchange occur in a leaf
§ They allow for the diffusion of oxygen and carbon dioxide into and
out of the leaf
• The epidermis is often covered by a waxy layer called the cuticle
o This forms an impermeable barrier

Mesophyll tissue
 • These are formed by parenchyma cells which contain chloroplasts
o This is where photosynthesis occurs in the leaf
• Two types of mesophyll tissue are found in the leaf:
o Palisade mesophyll forms a layer beneath the upper epidermis and contain
many chloroplasts for maximum photosynthesis
o Spongy mesophyll contains large air spaces between the cells for gas
exchange to occur

Vascular tissue

• Vascular tissue is arranged in vascular bundles and is responsible for the transport of
substances around the plant
o Vascular bundles form the veins in leaves
o Xylem transports water and mineral ions from the roots to the leaves
o Phloem transports the products of photosynthesis from the leaves to other
parts of the plant

Structure of a Leaf Diagram

The structure of a leaf has distinct layers each with their own function

Adaptations for gas exchange

• The leaf has several adaptations that facilitate gas exchange

Leaf Adaptations for Gas Exchange Table

 Adaptation Function
Prevents gases and water vapour from leaving through the epidermis so that gas
Waxy cuticle exchange must occur through stomata. This allows gas exchange and water loss to be
controlled
Contain stomata for gas exchange. Most stomata are found in the lower epidermis
Epidermis
where the temperature is lower. This reduces water loss
Maintains a concentration gradient of gases between the air and spongy mesophyll
Air spaces
cells by allowing movement of gases
Spongy
Increases the surface area for gas exchange
mesophyll
Guard cells Control gas exchange and water loss by opening or closing stomata
Xylem vessels bring water to the leaf which is required for photosynthesis and
Veins transpiration. Photosynthesis requires carbon dioxide to diffuse into the leaf while
transpiration involves the loss of water vapour
Transpiration: Consequence of Gas Exchange
• The majority of photosynthesis takes place in the leaves of plants
o Some plants are able to carry out photosynthesis in the cells of their stems
• During photosynthesis, carbon dioxide is taken in by the leaf and oxygen is
released
o The pores in the epidermis of the leaf through which this gas exchange takes
place are known as stomata (singular stoma)
o The stomata need to be open all the time in order for gas exchange, and
therefore photosynthesis, to continue
• The problem for plants is that as the stomata open to allow gas exchange to
occur, water in the form of water vapour is also lost through the stomata
o This water loss is known as transpiration
o Most plants can use cells called guard cells to close their stomata in order
to reduce water loss, but this will also reduce gas exchange and therefore
their rate of photosynthesis
o Transpiration is the inevitable consequence of gas exchange in the leaf
• There are some advantages to the process of transpiration
o It provides a means of cooling the plant via evaporation
o The transpiration stream is helpful in the uptake of mineral ions
o The turgor pressure of the cells, due to the presence of water as it moves up
the plant, provides support to the leaves and to the stems of non-woody plants
§ Leaves with high turgor pressure do not wilt and therefore have an
increased surface area for photosynthesis

Transpiration in the Leaf Diagram

The loss of water vapour from leaves by evaporation through the stomata is unavoidable as
gas exchange for photosynthesis can only occur when the stomata are open

Factors affecting the rate of transpiration

• Air movement
o More air movement leads to increased rates of transpiration
§ The air outside a leaf usually contains a lower concentration of water
vapour than the air spaces inside a leaf, causing water vapour
to diffuse out of the leaf
§ When the air is relatively still, water molecules can accumulate just
outside the stomata, creating a local area of high humidity
§ Less water vapour will diffuse out into the air due to the reduced
concentration gradient
§ Air currents, or wind, can carry water molecules away from the leaf
surface, increasing the concentration gradient and causing more
water vapour to diffuse out
• Temperature
o Higher temperatures lead to higher rates of transpiration, up to a point at
which transpiration rates will slow
§ An increase in temperature results in an increase in the kinetic
energy of molecules
§ This increases the rate of transpiration as water molecules evaporate
out of the leaf at a faster rate
§ If the temperature gets too high the stomata close to prevent excess
water loss
§ This dramatically reduces the rate of transpiration
• Light intensity
o Higher light intensities will increase the rate of transpiration up to a point
at which transpiration rates will level off
§ Stomata close in the dark and their closure greatly reduces the rate of
transpiration
 § Stomata open when it is light to enable gas exchange for
photosynthesis; this increases the rate of transpiration
§ Once the stomata are all open any increase in light intensity has no
effect on the rate of transpiration
• Humidity
o Higher humidity levels reduce the rate of transpiration
§ If the humidity is high that means the air surrounding the leaf
surface is saturated with water vapour
§ This causes the rate of transpiration to decrease as there is no
concentration gradient between the inside of the leaf and the outside
• At a certain level of humidity, an equilibrium is reached; water
vapour levels inside and outside the leaf are the same, so there
is no net loss of water vapour from the leaves

Several environmental factors affect the rate of transpiration in plants

Measuring the rate of transpiration

• The effect of environmental factors on the rate of transpiration in plants can be

measured using a piece of equipment called a potometer
o Note that while potometers are used to measure transpiration rates,
they technically measure the rate of water uptake rather than the rate of
transpiration, as a small amount of the water taken up by a plant will be used
in photosynthesis
§ Because the amount of water used in photosynthesis is so small in
relation to the total amount of water that passes through a plant,
 the rate of water uptake can reasonably be used to represent the
rate of transpiration
• Different types of potometer exist
o Bubble potometers measure the movement of an air bubble along a water-
filled tube connected to a plant shoot as water is drawn up by the shoot
§ The position of the air bubble is recorded at the start of an
experiment, and then a researcher can either measure how far the
bubble moves in a set amount of time, or time how long it takes for
the bubble to move a certain distance
o Mass potometers measure the change in mass of a water-filled test
tubeconnected to a plant shoot as it loses water over a set amount of time
• The effect of various environmental factors on transpiration can be measured by
placing the potometer in different conditions e.g.
o Wind speed
o Humidity
o Light intensity
o Temperature

A bubble potometer uses the movement of an air bubble to measure the rate at which water
is drawn up by a plant shoot. In this image the air bubble will move to the left along the
tube as the plant transpires

• Environmental factors can be investigated in the following ways

o Air movement
§ A fan on different settings could be used to vary the flow of air
around a plant shoot
o Humidity
 § Enclosing the plant shoot in a plastic bag can increase the humidity
§ A humidifier or dehumidifier could be used to give a measurable
variation in humidiy
o Light intensity
§ A lamp at different distances or with different types of light
bulb can be used to vary light intensity
o Temperature
§ A thermometer or temperature probe can be used to find
surroundings with different air temperatures
§ A heater or air conditioner can be used to give a measurable
variation in temperature
• A researcher would need to be aware of the importance of controlling any variables
other than the variable being investigated to ensure that any results are valid e.g.
placing a plant shoot in different rooms could be a way of varying temperature, but
might bring the risk of also varying light levels and humidity; these variables would
need to be controlled

Drawing Leaf Structure

• You will be expected to identify the following structures in the leaf of a
dicotyledonous plant:
o Chloroplasts
o Cuticle
o Guard cells
o Stomata
o Upper and lower epidermis
o Palisade mesophyll
o Spongy mesophyll
o Air spaces
o Vascular bundles (xylem and phloem)

Structure of Leaf Diagram

 Diagram showing the transverse section of a leaf

Drawing a plan diagram

 • Plan diagrams are drawings made from micrographs or from viewing specimens
under a low magnification
• Keep the following in mind when drawing a plan diagram:
o No individual cells are drawn, only tissue layers enclosed by lines should be
present
o Pay attention to the distribution of tissue throughout the plant organ
o Use a sharp pencil and draw clear, continuous lines
o Do not shade any part of your drawing
o Make sure your proportions and observations are accurate
§ Draw what you actually see, not what you would expect to see from a
textbook
o Draw your drawing big enough to fill up at least half the available space
• When labelling your plan diagram remember to:
o Use a ruler to draw label lines, not freehand
o Avoid using arrowheads and make sure the label lines stop at the structure
o Make sure label lines do not cross each other
o Write all labels horizontally, not at different angles

Worked Example

The following micrograph shows a transverse section of a dicotyledonous leaf.

Draw a labelled plan diagram of this micrograph.

By Berkshire Community College Bioscience Image Library, Public domain, Wikimedia

Answer:
Determining Stomatal Density
• The density of stomata (the number of stomata per unit of area) can be a useful
measurement to biologists
o To assess the plant's likely response to a dry spell of weather
o To predict its behaviour in windy or wet climates if the plant was being
moved for agricultural / horticultural reasons
• This technique can be used to assess how stomatal density varies from species to
species

Apparatus

• A plant to sample a leaf from

• Clear nail varnish (ideally solvent based)
• Sellotape
• Microscope
• Microscope slides
• Stage micrometer
• Counting device (clicker/ phone app etc.)
• Calculator

Method

• Select a leaf from a live plant and cut it off the plant
o Geraniums and spider plants make good subjects for this experiment
• Place the leaf upside down on a flat surface such as a tile or worktop
• Paint clear nail varnish onto the underside of the leaf
• Wait for the nail varnish to dry (approx. 5 minutes)
• Peel off the layer of varnish using sellotape
o Discard the leaf
o The layer of varnish now forms a leaf cast
• Place the dried varnish impression on a microscope slide
o A coverslip is not required as this isn't a biological sample, just an
impression of one
o A drop of water is not required either, so long as the sample is laid flat on the
slide
 • Use the usual steps to focus on the sample (you can read about this in our revision
note on microscope skills)
• Adjust the zoom such that a countable number of stomata are visible in the field of
view
o Between 15 and 100 is ideal
o Even if a stoma is partially visible at the edge, still count it as 1
• Count the stomata in that field of view
o You may wish to use a clicker or phone app so you don't lose count!
• Move the field of view to another area of the nail varnish layer and repeat
• Count at least 3 separate fields of view and take a mean value
o Repeat readings allow you to eliminate anomalous results and calculate
a reliable mean

Measurements to take

• Use a stage micrometer to measure the diameter of the field of view

o This has to be at the same magnification power that you used when counting
the stomata
• The stage micrometer will be calibrated in micrometers
o A typical microscope allows the scientist to look at a field of view of about 0.5
mm diameter when on full power (× 400)
• You will have calculated a mean number of stomata per field of view from the
previous stage
• You can read about using a stage micrometer in our revision notes on microscope
skills

Worked Example

A study reveals a mean count of 16 stomata per field of view at a magnification of × 400.
The stage micrometer calculates the diameter of the field of view at a magnification of × 400
to be 0.46mm

Calculate the stomatal density based on these data. Give units in stomata per mm2

Use a value of π = 3.14 and give your answer to the nearest whole number of stomata.
Answer:

Step 1: Calculate the radius of the field of view

Radius = Diameter ÷ 2

Radius = 0.46 mm ÷ 2 = 0.23 mm

Step 2: Calculate the area of the field of view

Area = πr2 = π × 0.232

Area = 0.1662 mm2

Step 3: Divide the mean number of stomata by the area of the field of view to calculate
density

Density = 16 ÷ 0.1662 = 96.27 stomata per mm2

Step 4: Round to the required precision (nearest whole number)

Density = 96 stomata per mm2

Limitations

• Not all plant species have easily accessible stomata that create a strong imprint
• Solvent-based nail varnish can destroy some of the cell structure it comes into contact
with
• Does the plant grow more stomata (guard cells) according to the conditions in each
individual habitat?
• Water-based nail varnish is safer to use but dries more slowly

NOS: Reliability of quantitative data is increased by repeating measurements

• Reliability refers to the level of trust that we can place in numerical measurements
o These types of measurements are known as quantitative data
• Repeating the stomatal count for the same species of leaf under the same conditions
will increase the reliability of the results
o It is very possible that the data gathered during a single count could contain
errors that we may not be aware of
o Taking repeated measurements will identify anomalous measurements and
allow us to calculate a mean
§ Anomalous measurements are those that deviate from the expected
measurements
§ Anomalies are omitted when calculating the mean for a data set
• If repeated stomatal counts deliver similar results, the data is said to be reliable
o We can therefore place a higher level of trust in the data than what would have
been possible if we got very different results with every count
• Repeating measurements is a crucial step in gathering data during a scientific
investigation