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1 cm sodium
carbonate solution
0.5 cm sodium
carbonate solution
2 cm acetic acid
4 cm acetic acid
Place 5 cm3 starch solution in each tube. Note the time and add 1 cm3 amylase solution to each
Then test for starch using iodine take a sample every 30 seconds and time how long it takes for the
iodine to no longer turn blue/black.
c. To demonstrate osmosis, Visking tubing (dialysis tubing) can be tied at one end and filled with 20 per
cent sucrose solution. The other end is attached to a capillary tube. The level of the sucrose can be noted
before and after the tubing has been placed in a beaker of water for about 30 minutes.Visking tubing has
microscopic holes in it, which let small molecules like water pass through (it is permeable to them) but is
not permeable to some larger molecules, such as the sugar sucrose. This is why it is called partially
permeable.
The sucrose molecules are too big to pass through the holes in the partially permeable membrane. The
water molecules can pass through the membrane in either direction, but those on the right are attracted
to the sugar molecules. This slows them down and means that they are less free to move they have less
kinetic energy. As a result of this, more water molecules diffuse from left to right than from right to left.
In other words, there is a greater diffusion of water molecules from the more dilute solution (in this case
pure water) to the more concentrated solution.
d. Onion epidermis can be peeled away, cut into squares and mounted on slides in different
concentrations of sucrose solution. Observation under a microscope will show the effects of osmosis.
Plasmolysis - The state of a plant cell in a hypertonic solution, the cell shrinks.
Turgid - Plant cells in a hypotonic solution, the cell swells against the cell wall and the cell is rigid and firm
Flaccid - Plant cells in a hypertonic solution, the cell shrinks away from the cell wall and the cell is limp.
e. Red blood cells in blood obtained from a butcher may be mounted on slides in hypotonic, isotonic and
hypertonic saline, and observed under a microscope to show the effects of osmosis.
Blood plasma has a concentration equivalent to a 0.85% salt solution. If fresh blood is placed into
solutions with different concentrations, the blood cells will gain or lose water by osmosis. This can be
demonstrated using sterile animal blood
(available from suppliers of biological materials).
Three test tubes are set up, containing these solutions:
A 10 cm of distilled water (hypotonic)
B 10 cm of 0.85% salt solution (isotonic)
C 10 cm of 3% salt solution (hypertonic)
1 cm of blood is added to each tube, and the tubes are shaken. A sample from each tube is examined
under the microscope. The sample from tube A is found to contain no intact cells
The blood cells on the right (tube C) were placed in a 3% salt solution and the normal blood cells on the
left(tube B) were in a 0.85% salt solution.
The cells from tube B look normal, but those from tube C are shrunken, with crinkly edges
It is important that animal cells are surrounded by a solution containing the correct concentration of
dissolved solutes. If the surrounding solution does not have the right concentration, cells can be damaged
by the effects of osmosis. The red blood cells placed in water absorb the water by osmosis, swell up and
burst, leaving a red solution of haemoglobin in the test tube. When placed in 3% salt solution, the red
blood cells lose water by osmosis and shrink.
The three tubes are now placed in a centrifuge and spun around at high speed to separate any solid
particles from solution.
Tube A contains a clear red solution and no solid material at the bottom of the tube = the cells have burst
due to water entering via osmosis and haemoglobin is released into the water
Tubes B and C both contain a colourless liquid and a red precipitate at the bottom = the cells have
remained intact so haemoglobin remains in the cells.
f. Osmosis can be demonstrated by using strips of potato, and this basic experimental method provides a
good opportunity for students to carry out individual whole investigations.
Because of the difficulty of the osmosis concept, it is better to keep this investigation until the latter part
of the course so that students will have had previous experience of carrying out investigations on simpler
topics. Students enjoy the reference to chips, but should quickly realise that it can be difficult to keep
the size constant to achieve consistency lengths of potato tissue can be drilled from a potato using a
cork borer. The chips are measured by mass or by length and are placed into sucrose solutions of
different concentrations for at least one hour. The percentage change in mass or length is a measure of
the degree of osmosis that has occurred.
A potato tuber is a plant storage organ. It is a convenient tissue to
use to investigate the effects of osmosis on the mass of the
tissue.
A boiling tube is half-filled with tap water and a second with
concentrated (Molar) sucrose solution. A third tube is left empty.
A potato is chipped cut into chips 5cm x 1cm x 1cm, making
these measurements as accurate as possible to ensure the chips
have the same surface area. No skin is left on the potato (it would
provide a waterproof layer).
Each chip is gently blotted to remove excess moisture and
weighed to find the starting mass of each. One chip is placed into each of the three tubes, and after a
fixed time the chips are removed, gently blotted to remove excess moisture and reweighed. The chips
can be felt to see and change in texture and re-measured to see if there is any changed in dimensions
(length/width/breadth).
The change in mass (+/-) is calculated for each chip and the % change is found. (% change in mass takes
into account the difference in starting masses of the chip so they can be compared).
% change = change in mass x 100
Starting mass
Boiling tube with water - water moves into the cells
The water in the tube is more concentrated than the water in the cells, so water molecules will move
from an area of higher concentration of water (in the tube) to an area of lower concentration of water (in
the potato cells).
The cell membrane is semi-permeable so the water molecules can get through and each cell will swell as
the water flows in.
As all the cells swell, the potato chip will increase in mass, and length/width/breadth and become more
rigid.
Boiling tube with concentrated sugar solution - water moves out of the cells
The water in the tube is a lower concentration than the water in the cells, so water molecules will move
from an area of higher concentration of water (in the potato cells) to an area of lower concentration of
water (in the tube).
The cell membrane is semi-permeable so the water molecules can get through and each cell will shrink as
the water flows out.
As all the cells shrink, the potato chip will decrease in mass, and length/width/breadth and become more
flexible/floppy.
Boiling tube with nothing - some water moves out of the cells
Water molecules will move from an area of higher concentration of water (in the potato cells) to an area
of lower concentration of water (in the tube).
The cell membrane is semi-permeable so the water molecules can get through and each cell will shrink as
the water flows out.
As all the cells shrink, the potato chip will decrease in mass, and length/width/breadth and become more
flexible/floppy. The effect will be less than in the tube with concentrated sugar solution.
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g. A variation on this theme is to cut potato cubes of different sizes, which have different surface area to
volume ratios. After measuring and recording the masses of the cubes, they are immersed in water. After
one hour, the cubes are blotted dry and their masses measured and recorded again. The percentage
increase in mass for cubes of different surface area to mass ratio can be compared, to explore the
concept of how surface area to volume ratio influences water uptake.
The surface area over which the osmosis occurs can change the speed at which it happens dramatically.
The larger the surface area between the two solutions the faster osmosis will happen. This is due to more
of each solution will be in contact with the membrane at one moment. This
means that a larger amount of the free water molecules in the solution can move across the membrane
at the same time therefore making the process happen much faster.
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Re-light a glowing
splint
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b. To measure the rate of oxygen production, the stem of a water plant is cut under water, andthe plant
kept immersed in water in a beaker or boiling tube. The number of bubbles of gasgiven off over a
measured time period can be counted. This simple experimental set up canenable students to carry out
individual investigations into the effect of different factors onthe rate of bubble production. Suitable
variables include: light intensity (the plant is exposedto a light source and the rate of bubble production
measured at different light intensities bychanging the distance between the light source and the water
plant); colour/wavelength oflight (coloured filters are placed between the plant and the light source); and
carbon dioxideavailability (the plant is immersed in solutions of different concentration of
sodiumhydrogen carbonate).
It is vital to have a stabilising period of a minimum 5 minutes before any measurements are taken.
An increase in light intensity should bring about an increase in gas production and, until a limiting factor
comes into operation, the two should be proportional.
Increasing the light intensity makes available a greater amount of energy to drive the photosynthetic
reaction. Since oxygen is a product of photosynthesis, a higher rate of photosynthesis should result in a
higher rate of oxygen production.
Unless the gas produced by Elodea is pure oxygen it would be possible for the plant to double its oxygen
output without doubling the volume of gas if the proportion of oxygen in the gas mixture were increased.
One would need to know that the percentage of oxygen in the bubbles remains constant, before it could
be assumed that doubling the volume of gas meant doubling the production of oxygen.
It is quite likely that the gas production will increase in direct proportion to the light intensity over the
whole range of this experiment. Limiting factors may operate, however, at the higher light intensities.
It is unlikely that gas production would continue to increase indefinitely with increasing light intensity
since carbon dioxide would become limiting or internal factors such as the available chloroplasts or the
diffusion rate of carbon dioxide would impose a limit on the rate of photosynthesis.
As the bench lamp comes closer to the beaker it might be expected that there would be a rise in
temperature of the water in the beaker. Since a rise in temperature speeds up many chemical reactions it
might also accelerate some stages of photosynthesis. (In practice, the temperature is unlikely to rise more
than 1 C.)
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c. Starch production can be investigated by placing a plant in the dark for 24 hours to de-starch the
leaves. A starch test on a leaf from a plant that has been kept in the dark will not give a blue-black colour,
whereas a similar test on a control leaf from a plant kept in the light will give a blue-black colour.
Remove a green leaf from a plant that has been exposed to sunlight for a few hours
Half-fill a beaker with water. Heat the water until it boils. Keep the water at boiling point.
Use the forceps to place the leaf in the boiling water. Boil for 2 minutes.
Boiling the leaf in water:
Removes the waxy cuticle which prevents entry of iodine/potassium iodide solution.
Ruptures cell membranes to make starch granules in cytoplasm and chloroplasts accessible to
iodine/potassium iodide solution. Cell membranes are selectively permeable and do not readily
allow the penetration of iodine.
Denatures enzymes, particularly those which convert starch to glucose e.g. diastase.
Boiling arrests all chemical reactions, since enzymes which catalyse the reactions are denatured.
Denatured enzymes have altered or destroyed active sites due to heat, pH, ionic concentration
Place the boiled leaf in a boiling tube containing 90% ethanol.
Place the boiling tube in hot water and boil for 10 minutes or until the leaf decolourizes.
Boiling the leaf in ethanol:
Removes chlorophyll which is a green pigment and so masks the colour change of the iodine test
for starch, so the leaf needs to be ' decolourized' for changes to be observed. A decolourized leaf is
pale yellow or green. Ethanol is an organic solvent and so extracts chlorophyll from the leaf.
Gently remove the leaf and wash with a fine trickle of cold tap water.
Spread the leaf evenly on a white tile.
Add a few drops of iodine/potassium iodide solution to the leaf and note any observations.
The iodine solution will turn from brown to blue - black if starch is present.
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d. A starch test on a variegated leaf can be used to demonstrate that chlorophyll is needed for
photosynthesis.
Place a plant with variegated leaves in the dark for at least 24 hours to make sure that there is no
starch in the leaves i.e. de-starching the leaves.
Variegated Leaves: have green and white areas the green areas have chlorophyll but chlorophyll is
absent in the white areas.
Leave the plant in the light with de-starched leaves in good light for 6 hours at room temperature
(20C).
Then test the leaf for the presence of starch using iodine.
Compare the stain pattern with the colour map.
The green chlorophyll rich parts are blue-black with starch.
The white parts without chlorophyll are yellow-brown without starch.
Since starch is only produced if chlorophyll is present, then chlorophyll is needed for photosynthesis
e. To show that carbon dioxide is needed for photosynthesis, a leaf on a plant may be surrounded by air
with no carbon dioxide by inserting it into a conical flask containing a small amount of potassium or
sodium hydroxide. The plant is left in good light for 24 hours.
NaOH removes
CO
The test leaf and a control leaf from the plant are then tested for starch.
To test leaves for starch:
drop into very hot/boiling water for one minute (to destroy the cell membranes so that chlorophyll
molecules can pass through)
drop into hot ethanol (to remove/dissolve the green chlorophyll)
place leaf into water (to rehydrate and soften the leaf so that it can be spread out)
drop iodine solution onto the leaf (test for starch) blue-black colour will show the presence of starch.
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Bomb calorimeter
NB: The energy gets lost to the surroundings as not all energy is direct to heat the water, also there is a
need to heat the glass apparatus first.
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Vacuum flasks are needed for this activity. Surface sterilised seeds are put into a flask, which is sealed
with a bung. To show heat production the flask needs a cotton wool bung with a thermometer going
through the bung into the flask.
Students could demonstrate that they produce carbon dioxide by breathing out through a tube into lime
water or hydrogen carbonate indicator.
thermometer
cotton wool
Living seeds
Dead seeds
The temperature will increase significantly in the flask containing the live seeds. Respiration produces the
energy the seeds need to germinate and the by-product of the use of the energy is heat (energy).
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A glass tube runs from inside the flask, through the bung and into an indicator solution of either limewater
or hydrogen carbonate. The carbon dioxide produces changes the colour of the indicator solution.
The lime water associated with living organisms should be milky. The lime water receiving air from nonliving organisms should be clear.
Carbon dioxide is the gas which turns lime water milky.
The results suggest that carbon dioxide is given out by the organisms.
The air from non-living organisms failed to turn lime water milky. Thus the carbon dioxide which was in
sufficient concentration from living organisms to turn lime water milky so must have come from the living
organisms rather than the air around them
NB: The experiment provides evidence for only the organisms in the test-tubes.
The failure of dead organisms to turn lime water milky shows that the production of carbon dioxide results
from a process that occurs in living organisms but not in dead ones.
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7. Simple controlled experiments to investigate the effect of light on net gas exchange
from a leaf, using hydrogen carbonate indicator
A water plant can be placed in a sealed tube of air-equilibrated hydrogen carbonate solution (red in
colour) and placed in the light
or in the dark. The solution will
turn purple if kept in the light
and will turn yellow if kept in
the dark.
A variation could involve the
use of water snails or, if not
available, small land insects
placed on a gauze platform
above the indicator, with and
without the water plant. This
variation allows students to
think about the balance
between carbon dioxide used
by photosynthesis and carbon
dioxide produced by
respiration.
Tube
A
B
C
Colour of indicator
yellow
purple
red
A useful demonstration uses four tubes containing hydrogen carbonate solution: one with water plant
only, one with animals only, one with both water plant and animals and one with no living organisms.
One set of the tubes is exposed to light and left for 12 to 24 hours and another set is placed in the dark
for the same length of time.
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The breathing rate can be measured at rest and after a period of exercise by counting the number of
inhalations per minute.
Exercise also influences the rate of breathing by increasing the volume of each breath, this can be
measured by taking the volume of one exhalation before and after exercise.
This can be done by breathing through a tube into a plastic container filled with water. The volume of
displaced water can be measured. The breathing rate at rest and after exercise can be calculated as
number of breaths per minute x volume of each breath.
Volume of each
breath before
exercise
Volume of each
breath during exercise
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10. A simple experiment to investigate the effect of exercise on heart rate in humans
Students should be shown how to measure and record their pulse. They can measure this at rest.
A short period of exercise can follow, stepping on and off a low stool, or running up
and down a flight of stairs for five minutes. (Safety note: careful supervision is
required.)
The pulse rate should now be recorded for five minutes or until it returns to its rest
level. This experiment can provide a great deal of interesting data on the variation in
resting heart rate, the effects of exercise and how quickly the heart recovers.
Discussion can include measures of fitness, heart disease, cardiac output and the
effects of long-term exercise on stroke volume.
To get rid of the extra CO2 you start to breathe deeper and faster
Your heart rate increases to pump more oxygen around the body
Blood is diverted away from inactive organs (stomach / liver) and towards the working muscles through
vasoconstriction and vasodilation
When the muscles contract they squeeze blood back towards your heart
The heart contracts even stronger to pump more blood with each beat
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11. A simple experiment to show how the sensitivity of the skin differs on finger tips,
back of hand, wrist and forearm
Students should work in pairs. A piece of hard cardboard or cork can be used to fix the two prongs of a
hairpin or two pins 5 mm apart. This is then used by one
student to lightly touch the fingertips of another who is
looking away. The first student can use both points or one
point as a stimulus. The second student then has to judge
whether one or two points were used and their response
recorded as correct or incorrect. This can be repeated 10
times for each area of the hand. It is then repeated using
pins 1 cm apart and 2 cm apart.
Students can then identify the most sensitive area as this
should have the most correct responses with the smaller
distance. Conclusions can be made about the number of
sensory nerve endings, receptive field size and the
thickness of skin. This practical also provides
opportunities to discuss data analysis, experiment design
and anomalous results, and the benefits of grouping class
results.
Finger-tips will probably recognize the two points even when they are only about 2 mm apart,
i.e. the lower limit of this instrument. The back of the hand will probably need a separation of l0 mm to
achieve a score of 8-10. Other areas will probably be even less sensitive.
Possibilities:
(a) One point does not touch a receptor. It seems unlikely that, on the back of the hand, a pressure
actually indenting the skin will not affect at least one receptor. Moreover, recognition of two stimuli
when they are separated by half a second suggests that it is not shortage of receptors alone which causes
poor discrimination.
(b) Both points touch receptors which feed impulses into one nerve fibre. Since a single sensory fibre may
receive branches from sensory endings covering a region of several square millimetres this explanation is
quite plausible. The recognition of the non-simultaneous double stimulus could be explained by the
additional burst of impulses initiated by a second, stimulus. It has been claimed that single hair follicle
receptors are sensitive over an area several centimetres in diameter but the 65 mm separation claimed
for the back could hardly be explained in this way.
(c) There are two receptors and two sensory fibres but unless the impulses are separated in time, they
cannot be discriminated by the brain.
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12. A practical exercise comparing floral structure in insect-pollinated and windpollinated flowers
Most areas should have access to suitable specimens. Insect-pollinated flowers can be examined and the
various structures observed. Wind-pollinated grasses should be available but have fewer structures to
see.
Insect Pollinated
Wind Pollinated
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Plant material such as wheat, maize, oat or cress seedlings can be used to demonstrate phototropism.
Petri dishes containing moist cotton wool and the plant material can be put into light-proof boxes such as
shoe boxes. To create unilateral light a small slit can be cut in one side of the box and light can be shone
into the box. Control seedlings can either have aluminium caps put on their tips or can be kept in a shoe
box without a slit for light. A clinostat needs to be used to demonstrate geotropism.
To test the response to or effect of light on a plant shoot.
A growing herbaceous stem is placed in a box that has a small opening on one side only, where light
enters.
Auxins are plant hormones that make some parts of a plant stem grow faster than others. The result is
that the plant stem bends towards the light.
You may have noticed that a houseplant grows towards the window and turns its leaves towards the
light. It does this because light coming from the window side of the plant destroys the auxin in that side
of the stem. So growth on that side slows down.
On the shaded side of the plant there is more auxin. So growth on this side speeds up. The result is that
the shoots and leaves are turned towards the light for photosynthesis.
A few beans are soaked in water overnight then placed on wet cotton wool for a few days until the first
root of each seed (the radicle) grows to a length of about 2cm. Wet cotton wool is attached to the cork
disc of two clinostats. Three or four of the germinating bean
seeds are pinned onto each of the discs with their radicles
pointing outwards. Covers are placed on them to keep the
air moist. The clinostats are turned on their sides. One is left
switched on, the other is switched off to act as the control
(to allow you to compare your results) Both clinostats are
left set up for a few days. The cotton wool is kept damp.
After this time the radicles of the beans on the control
clinostat will have grown downwards, whereas those on
the beans on the rotating clinostat will be growing straight
out horizontally.
Explanation: Gravity would have been 'pulling' consistently
at 90 to the horizontal radicles in the stationary box.
In the clinostat the gravitational force also acts at 90 but on all sides of the radicle in turn.
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14. The use of quadrats to estimate the population size of an organism in two different
areas
Quadrats can be used to sample part of each area. Calculation will be needed to work out the estimated
population size. For example, if 10 quadrats have been used and the total area amounts to 100 quadrats,
the estimated population size will be the number of organisms counted in the 10 sample quadrats
multiplied by 10.
It isimportant to place the sample quadrats randomly this avoids bias and improves the reliability of the
data.
An interesting way to practise the technique is to throw plastic beads on the floor of the classroom and
ask students to guess how many beads there are. The quadrat sampling procedure can be used in front of
students to get an estimate. The beads can then be collected and counted.
The actual number can be compared to the estimated number and used to see how accurate the
estimation was.
A quadrat is a square (of either metal,
wood, or plastic) used in ecology and
geography to isolate a sample, usually
about 1m2 or 0.25m2. The quadrat is
suitable for samplingplants, slowmoving animals (such as millipedes
and insects), and some aquatic
organisms.
When an ecologist wants to know
how many organisms there are in a
particular habitat, it would not be
feasible to count them all. Instead, he
or she would be forced to count a
smaller representative part of the
population, called a sample. Sampling
of plants or animals that do not move
much (such as snails), can be done
using a sampling square called a
quadrat. A suitable size of a quadrat
depends on the size of the organisms
being sampled. For example, to count
plants growing on a school field, one
could use a quadrat with sides 0.5 or
1 metre in length.
It is important that sampling in an area is carried out at random, to avoid bias. For example, if one were
sampling from a school field, but for convenience only placed quadrats next to a path, this might not give
a sample that was representative of the whole field. It would be an unrepresentative, or biased, sample.
One way one can sample randomly is to place the quadrats at coordinates on a numbered grid.
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This allows students to see that carbon dioxide is released during anaerobic respiration (fermentation).
Students add yeast to glucose solution in a side-arm test tube. Anaerobic conditions are achieved by
putting a drop of oil (cooking oil will do) onto the yeast and glucose mixture. A rubber tube is attached to
the side arm of the test tube and a glass pipette is inserted at the other end of the rubber tube. The
pipette is placed under water to allow the bubbles of carbon dioxide gas to be counted. Temperature is
the easiest condition to investigate. Glucose concentration and pH could also be investigated.
tap open
3-way tap
(open)
capillary
tube
water bath
tap closed
yeast and
glucose A
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