Chapter 4 Cell Membrane and Transport

4.1 Fluid Mosaic Membranes

4.2 Movement into and out of Cells

4.1 Fluid Mosaic Membranes

1) The Fluid Mosaic Model

 Characteristics of Membranes
• partially permeable
• about 7nm wide
• formed from a bilayer of phospholipids
 Function of Membranes
• The cell surface membrane separates the internal cell environment from the
external environment
• The cell surface membrane acts as a barrier to water soluble molecules.
• Intracellular membranes form compartments within the cell.
• Membranes control the exchange of materials across them
• Membranes contains protein receptors and acts as an interface for communication
(cell signalling, cell recognition)

Phospholipids
 Structure of Phospholipid
• phosphate head (polar; hydrophilic)
• fatty acids tail (non-polar; hydrophobic)
 If phospholipid molecules are spread out over the surface of water,
they form a phospholipid monolayer.
• a single layer with the hydrophilic phosphate heads in the water and the
hydrophobic fatty acid tails project out of the water.

 If phospholipids are mixed with water,

they form a micelle (sphere shape).
• In a micelle, the hydrophilic phosphate heads facing out towards the water and the
hydrophobic fatty acid tails facing in towards each other, creating a hydrophobic
environment.
or, they form phospholipid bilayers (sheet shape).
• the basic structure of membranes
• can form compartments

The Fluid Mosaic Model

Fluid - the phospholipids and proteins can move around the monolayer by diffusion.
- the phospholipids mainly move sideways, within their own layers
(*they do not flip to the other monolayer)
- the membrane is dynamic

Mosaic - proteins' random arrangement on the surface of the phospholipid bilayer.

- describes the pattern produced by proteins scattered in the phospholipid
bilayer.
- there are many types of proteins (transport proteins, receptors, enzymes, etc.)
 2) Components of Cell Surface Membranes
3 types of lipids 2 types of proteins
- phospholipids - glycoprotein (with polysaccharides)
- cholesterol - other proteins
- glycolipid (with polysaccharides)

①) Phospholipids
→ Form a bilayer (two layers of phospholipid molecules).
→ Hydrophobic tails (fatty acid chains) point in towards the membrane interior.
→ Hydrophilic heads (phosphate groups) point out towards the membrane surface.
→ Individual phospholipid molecules can move around within their own monolayers by
diffusion.
→ Some of the fatty acids tails are saturated and some are saturated.

②) Cholesterol
→ Cholesterol molecules also have hydrophobic tails(non-polar region) and hydrophilic
heads (polar region)
→ Fit between phospholipid molecules and orientated the same way (head out, tail in)
→ Cell surface membranes in animal cells contain almost as much cholesterol as
phospholipid.
→ Cholesterol is much less common in plant cell membranes.
→ Are absent in prokaryotes membranes

③) Glycolipids
→ Lipids with carbohydrate chains attached
→ These carbohydrate chains project out into the fluid surrounding the cell (they are
found on the outer phospholipid monolayer).

④) Glycoproteins
→ Proteins with carbohydrate chains attached
→ These carbohydrate chains project out into the fluid surrounding the cell (they are
found on the outer phospholipid monolayer)

The carbohydrate chains

• are short polysaccharides composed of a variety of different monosaccharides
• form a cell coat / glycocalyx outside the cell membrane.
→ Intrinsic Proteins (or Integral Proteins)
▪ The proteins embedded within the membrane.
▪ They can be in the inner or outer phospholipid monolayer
▪ Most commonly, they span the entire membrane – these are known as
transmembrane proteins. Eg: transport proteins.
→ Extrinsic Proteins (or Peripheral Proteins)
▪ Proteins that are found on the inner or outer surface of the membrane
→ Proteins can slide around the membrane but can never flip from one side to
the other.
→ Proteins have a polar region where hydrophilic amino acids in contact with the
water on the outside of the membranes.
Proteins have a non-polar region where hydrophobic amino acids in contact
with the fatty acid tails inside the membrane.
3) Roles of Molecules in the Cell Surface Membrane

(1) Phospholipids
1. Form the basic structure of the membrane (phospholipid bilayer)
2. Act as a barrier to most water-soluble substances
(The fatty acid tails form a hydrophobic core in the phospholipid bilayer.
The non-polar fatty acid tails prevent polar molecules or ions from passing across the
membrane)
• This ensures water-soluble molecules such as sugars, amino acids and proteins
cannot leak out of the cell and unwanted water-soluble molecules cannot get in
3. Can be chemically modified to act as signalling molecules by:
• Moving within the bilayer to activate other molecules (e.g. enzymes)
• Being hydrolysed which releases smaller water-soluble molecules that bind to
specific receptors in the cytoplasm
4. Affects the fluidity of the membrane:
• the length of the fatty acids tails
• how saturated or unsaturated the fatty acid tails are

(2) Cholesterol
1. Cholesterol regulates the fluidity of the membrane
2. Cholesterol is important for the mechanical stability of membranes
• Cholesterol sit in between phospholipids molecules and reduce the fluidity of the
membrane which strengthens it.
• Prevent breakage and bursting.
3. Cholesterol contributes to the impermeability of the membrane to ions and polar
molecules.
(The hydrophobic regions of cholesterol molecules help to prevent ions or polar
molecules from passing through the membrane.
• The myelin sheath which surrounds nerve cells is made up of many layers of cell
surface membranes to prevent leakage of ions which would slows down nerve
impulses.
4. At low temperature, cholesterol prevents membranes from freezing and fracturing.
• Cholesterol molecules sit in between the phospholipids, cholesterol prevents
phospholipids from packing too closely together when the temperature is low.
Membranes will not freeze easily.
5. At high temperature, cholesterol stabilises the cell membrane by stopping the membrane
from becoming too fluid.
• Cholesterol molecules bind to the hydrophobic tails of phospholipids, stabilising
them and causing phospholipids to pack more closely together.
The membrane becomes less fluid.
(3) Glycolipids and Glycoproteins
1. Acts as receptor molecules
• The carbohydrate chains enable the glycoproteins and glycolipids to acts as
receptor molecules.
• This allows glycolipids and glycoproteins to bind with certain substances at the
cell’s surface
• There are three main receptor types:
→ signalling receptors for hormones and neurotransmitters
→ receptors involved in endocytosis
→ receptors involved in cell adhesion and stabilisation
(as the carbohydrate part can form hydrogen bonds with water molecules
surrounding the cell)
2. Cell-to-cell recognition
Some act as cell markers or antigens, allowing cells to recognize each other.
• The carbohydrate chains bind to complementary sites on other sites.
• e.g. the ABO blood group antigens are glycolipids and glycoproteins that differ
slightly in their carbohydrate chains.

3. Stabilizing the Cell Surface Membrane

• glycoproteins and glycolipids form hydrogen bonds with water molecules both
inside and outside the cell surface membrane.

(4) Proteins
1. Stabilizing the membrane
• The interaction between the polar region of the protein with water on the
surface of the membrane and the interaction between the non-polar region of
the protein with fatty acids chains inside the membrane helps maintain the
stability of the membrane.
2. Transport Proteins
• Transport proteins provide hydrophilic channels for ions and polar molecules to
pass through the membrane.
• Each transport protein is specific for a particular kind of ion or molecule.
• 2 types of transport protein
→ Channel Proteins
→ Carrier Proteins
• Transport proteins allow the cell to control which substances enter or leave.
3. Enzymes
• Digestives enzymes found in the cell surface membranes of the cells lining the
small intestines.
• Enzymes involved in cellular respiration(in mitochondria) and
photosynthesis(chloroplasts)
 4. Cytoskeleton
• Some proteins on the inside of the cell surface membrane are attached
to the cytoskeleton.
• These proteins help to maintain and decide the shape of the cell.
• They may be involved in changes of shape when cells move.

Membranes become less fluid when there is:

• An increased proportion of saturated fatty acid chains,
As these chains are straight,
the chains pack together tightly
there are more intermolecular forces between the chains.
Membrane fluidity decreases

• A lower temperature ,
the molecules have less energy
therefore they are not moving as freely which causes the structure to
be more closely packed
Membrane fluidity decreases.

• An increase in the number of cholesterol molecules

Membranes become more fluid when there is:

• An increased proportion of unsaturated fatty acid chains,
As these chains are bent,
which means the chains are less tightly packed together,
there are less intermolecular forces between the chains.
Membrane fluidity increases.

• At higher temperatures,
the molecules have more energy ,
therefore they move more freely.
Membrane fluidity increases

• A decrease in the number of cholesterol molecules

 4) Cell Signalling
 Cell signalling is the process by which messages are sent to cells
 Cell signalling is very important as it allows multicellular organisms to control / coordinate
their bodies and respond to their environments.
 Stimuli can be from inside(hormones) and outside(light)
 Types of Signalling Pathway:
• electrical (nerves)
• chemical (hormones)
 Distance of Cell Signalling:
• Endocrine Signalling
- signalling over large distances, often through circulatory system.
• Paracrine Signalling
- signalling between cells close together, either through extracellular fluids or
directly between cells.
• Autocrine Signalling
- cell stimulates response within itself by releasing signals for its own receptors.
 Ligands: Signalling molecules.

First Part of a Cell Signalling Pathway

①) Ligands are secreted from a cell (the sending cell) into the extracellular space
②) The ligands are then transported through the extracellular space to the target cell
③) The ligands bind to surface receptors (specific to that ligand) on the target cell
- These receptors are formed from glycolipids and glycoproteins
④) The ligands bring about a change in the shape of the receptors.
⑤) Transduction: The signal is converted to a message that is readable.
⑥) Transmission of message to the effector.
⑦) The effector produces an appropriate response.
  Types of Signalling Molecules:
- hydrophobic signalling molecules (not water-soluble)
▪ Eg: steroid, hormones
- hydrophilic signalling molecules (water-soluble)

Second Part of a Cell Signalling Pathway

 Typical Signalling Pathway (for water insoluble signalling molecules)
①) Hydrophobic signalling molecules can diffuse directly through the cell surface membrane.
②) The signalling molecules bind to receptors in the cytoplasm or nucleus.
(Hydrophilic signal molecule)

 Typical Signalling Pathway (for water soluble signalling molecules)

1) Signal (ligand) arrives at a protein receptor in the cell surface membrane.
• Receptor is a specific shape which recognizes the signal.
• Only cells with this receptor can recognize the signal.

2) Signal changes the shape of the receptor, and since the receptor span the membrane, the
message is passed to the inside of the cell (signal transduction).
• Changing the shape of the receptor allows it to interact with the next component of the
signally pathway (in this case, G-proteins), so the message gets transmitted.

3) The next component is usually a G-protein which act as a switch mechanism to bring about
the release of a 'second messenger'.
The second messenger is a small molecule which diffuses through the cell relaying the
message.
• G-proteins are so-called because the switch mechanism involves binding to GTP
molecules (guanine triphosphate, similar to ATP).

4) Amplification occurs.
- After one receptor molecule is stimulated, many second messenger molecules will be
made in response .
- Second messenger usually activates an enzyme, which in turn activates further
- enzymes, increasing amplification at each stage.

5) Finally, enzymes are produced which bring about the required change in cell metabolism.
Response is produced.
Signalling cascade: The sequence of events including amplification triggered by G-protein.
Example of Cell-Signalling Pathway Involving a Second Messenger:
- Pathway involving the hormones glucagon and adrenaline

Apart from second messengers,

Other ways receptors can alter activity of the cell:
→ Opening an ion channel, resulting in change of membrane potential
- e.g. nicotine-accepting acetylcholine receptors

→ Acting directly as a membrane-bound enzyme

- e.g. glucagon receptor

→ Act as a intracellular (inside the cell) receptor when initial signal passes through
- e.g. the oestrogen receptor is in the nucleus and directly controls gene expression
when combined with oestrogen
4.2 Movement Into and Out of Cells
Types of Transport Mechanism:
 Diffusion
 Facilitated Diffusion
 Osmosis
 Active Transport
 Bulk Transport

1) Diffusion

Diffusion
the net movement of molecules or ions from a region of higher concentration to a region of
lower concentration, down a concentration gradient, as a result of the random movements of
particles.

 The particles move down a concentration gradient.

 The random movement is caused by the natural kinetic energy of the molecules or ions.
 As a result of diffusion, molecules or ions tend to reach an equilibrium situation where
they are evenly spread out within a given volume of space.

Particles that can pass through the cell surface membrane by diffusion:
- uncharged, non-polar molecules
Eg: O2 , CO2
- hydrophobic molecules
- small molecules
Eg: H2O (polar)

Factors that affect the rate of diffusion

①) Steepness of the concentration gradient
②) Temperature
③) The properties of the molecules or ions
- size of the molecules or ions
- polarity/solubility of the molecules or ions
④) Surface area of membrane
⑤) Diffusion distance (thickness of membrane)
⑥) Permeability of membrane
1) The steepness of the concentration gradient
→ The steepness of the gradient is the difference in the concentration of the substance on the
two sides of the membrane.

→ If there are more molecules of the substance on one side of the membrane than on the
other, there will be a net movement of molecules from where there are more to where
there are fewer

→ The greater the difference in concentration, the steeper the concentration gradient of a
substance across a membrane, the faster the rate of diffusion of that substance.

2) Temperature
At high temperatures, molecules and ions have high kinetic energy, they move faster, rate
of diffusion increases.

3) The properties of the molecules or ions

→ Size of particles
• Large molecules require more energy to get them moving than small molecules do, so
large molecules diffuse more slowly than small molecules.
→ Polarity of particles
• Uncharged and non-polar molecules diffuse directly across the phospholipid bilayer.
• Non-polar molecules diffuse more quickly than polar molecules because they are
soluble in the non-polar phospholipid bilayer.

4) Surface Area
→ The greater the surface area, the greater the number of molecules or ions can diffuse
across it per unit time, and therefore the rate of diffusion is faster.

→ The surface area of cell membranes can be increased by folding.

Eg: - microvilli in the cell linings the intestine
- cristae inside mitochondria

→ When the size increases, the volume increases faster than surface area, the surface area to
volume ratio decreases.

→ As the size of a cell increases, the surface area to volume ratio decreases, the lower the
number of molecules or ions can diffuse across it per unit time, the rate of diffusion
decreases.
 2) Facilitated Diffusion
Facilitated Diffusion
the transport of substances across a partially permeable membrane by a transport protein
molecule down a concentration gradient.

 Types of substances cannot diffuse through the phospholipid bilayer of cell membranes:
- Large polar molecules
Eg: glucose and amino acids
- Ions
Eg: sodium ions (Na+) and chloride ions (Cl-)

 These substances can only cross the phospholipid bilayer with the help of certain proteins

 There are two types of proteins that enable facilitated diffusion:

- Channel proteins
- Carrier proteins
 They are highly specific (they only allow one type of molecule or ion to pass through).

Channel Proteins
→ Channel proteins are water-filled pores.

→ They allow charged substances (eg. ions) to diffuse through the cell membrane.

→ The diffusion of these ions does not occur freely, most channel proteins are ‘gated’,
- part of the channel protein on the inside surface of the membrane can move to close
or open the pore.
- This allows the channel protein to control the exchange of ions.
Carrier Protein
→ Unlike channel proteins which have a fixed shape, carrier proteins can switch between
two shapes.

→ This causes the binding site of the carrier protein to be open to one side of the membrane
first, and then open to the other side of the membrane when the carrier protein switches
shape

Factors that Affect Rate of Diffusion through Channel and Carrier Proteins

→ The direction of movement of molecules diffusing across the membrane depends on their
relative concentration on each side of the membrane

The molecules will bind on the side where the molecule's concentration is high and be
released on the side where the molecule's concentration is low.

Net diffusion of molecules or ions into or out of a cell will occur down a concentration
gradient from a higher concentration to a lower concentration.

→ Factors:
- Steepness of the concentration gradient
(relative concentration on each side of the membrane)

- Number of transport proteins available in the membrane

(for channel protein, number of channel proteins that is open)

- Temperature
 3) Osmosis
Osmosis
the net movement of water molecules from a region of higher water potential (dilute solution)
to a region of lower water potential (concentrated solution) through a partially permeable
membrane.

 All cells are surrounded by a cell membrane which is partially permeable

 The cell membrane is partially permeable which means it allows small molecules (like
water) through but not larger molecules (like solute molecules)

 Water can move in and out of cells by osmosis

 Water is moving down its concentration gradient

Water Potential is a measure of the tendency of water to move out of a solution

 A dilute solution has a high water potential.

A concentrated solution has a low water potential.

 The water potential of pure water (without any solutes) at atmospheric pressure is 0kPa,
any solution that has solutes will have a water potential lower than 0kPa (a negative value).

Factors that affect water potential in osmosis:

1) the concentration of the solution
(the solute potential, s - the amount of dissolved solute present in a solution)

2) the amount of mechanical pressure applied to a solution

- a pressure greater than atmospheric pressure is applied to the solution
(the pressure potential, p - mechanical pressure acting on the solution)
= s + p
Osmosis in Animal Cells
Isotonic solution is a solution of equal water potential to a cell.
Hypertonic solution is a solution of lower water potential than a cell.
Hypotonic solution is a solution of higher water potential than a cell.

1) In a hypotonic solution/dilute solution/pure water,

• The dilute solution has a higher water potential, the cell has a lower water potential.
• Water will enter the cell through its partially permeable cell surface membrane by
osmosis.
• The cell will gain water by osmosis and it's volume increases
• The cell will continuously gain water until the cell membrane is stretched too far and the
cell bursts (cytolysis), as it has no cell wall to withstand the increased turgor pressure
created.

2) In an isotonic solution
• The isotonic solution has the same water potential as the cell.
• The movement of water molecules into and out of the cell occurs at the same rate
• No net movement of water
• No change to the cells

3) In a hypertonic solution/concentrated solution,

• The concentrated solution has a higher water potential, the cell has a lower water
potential.
• Water will leave the cell through its partially permeable cell surface membrane by osmosis
• The cell will lose water and it's volume decreases.
• The cell shrinks, shrivel up and crenates. (Crenation)
Osmosis in Plant Cells
1) In a hypotonic solution/dilute solution/pure water,
• The dilute solution has a higher water potential, the plant cell has a lower water potential.
• Water will enter the plant cell through its partially permeable cell surface membrane by
osmosis.
• As water enters the vacuole of the plant cell, the volume of the plant cell increases.
• The expanding protoplast pushes against the cell wall .
• Pressure builds up inside the cell
 This pressure increases the water potential of the cell until the water potential inside
the cell equals the water potential outside the cell, and equilibrium is reached.
• The inelastic cell wall prevents the cell from bursting
 The pressure created by the cell wall also stops too much water entering.
 The pressure builds up quickly, it takes very little water to enter the cell to achieve
equilibrium.

- When a plant cell is fully inflated with water and has become rigid and firm, it is described
as "fully turgid".
- Turgidity is important for plants as the effect of all cells in a plant being firm will provide
support and strength for the plant, making the plant stand upright with its leaves held out
to catch sunlight
- If plants do not receive enough water the cells cannot remain rigid and firm (turgid) and the
plant wilts.

2) In a hypertonic solution/concentrated solution

• The concentrated solution has a higher water potential, the plant cell has a lower water
potential.
• Water will leave the plant cell through its partially permeable cell surface membrane by
osmosis
• As water leaves the vacuole of the plant cell, the volume of the plant cell decreases.
• The protoplast gradually shrinks and no longer exerts pressure on the cell wall.
 Incipient plasmolysis is the point at which plasmolysis is about to occur .
 This is the point when the protoplast no longer exerts any pressure on the cell wall.

• As the protoplast continues to shrink, it begins to pull away from the cell wall
• This process is known as plasmolysis – the plant cell is plasmolysed.
 Plasmolysis - the loss of water from a plant or prokaryote cell to the point where the
protoplast shrinks away from the cell wall.

• External solution will pass through the cell wall and is still in contact with the protoplasm.
 4) Active Transport
Active Transport
the movement of molecules and ions through transport proteins across a cell membrane
against concentration gradient (from a region of lower concentration to a region of higher
concentration ) using energy from ATP.

• Active transport requires carrier proteins

- each carrier protein being specific for a particular type of molecule or ion.

• The energy is required to make the carrier protein change shape, allowing it to transfer the
molecules or ions across the cell membrane.

• The energy required is provided by ATP (adenosine triphosphate) produced during

respiration.

Processes involving active transport:

→ Nerve impulse transmission.
→ Muscle contraction.
→ Protein synthesis.
→ Excretion of urea by kidney.
→ Reabsorption of useful molecules and ions into the blood after filtration into the kidney
tubules.
→ Absorption of some products of digestion from the digestive tract.
→ Loading sugar from the photosynthesising cells of leaves into the phloem tissue for
transport around the plant.
→ Loading inorganic ions from the soil into root hairs.

Cells involved in active transport:

- Contain many mitochondria
- Have high rate of respiration

Sodium–potassium pump (Na+–K+ pump)

①) The role of the sodium–potassium pump is to pump 3 sodium ions out of the cell at the
same time as allowing 2 potassium ions into the cell for each ATP molecule used.

②) Sodium and potassium ions are both positively charged.

③) So the net result is that the inside of the cell becomes more negative than the outside.

④) A potential difference (p.d.) is created across the membrane.

 5) Endocytosis and Exocytosis
The processes of diffusion, osmosis and active transport are responsible for the transport of
individual molecules or ions across cell membranes.

Bulk transport is responsible for the transport of larger quantities of materials including large
molecules into or out of cells.

Materials that are transported via bulk transport:

→ Large molecules such as proteins or polysaccharides
→ Parts of cells
→ Whole cells eg. bacteria

Bulk Transport
→ Endocytosis - bulk transport into cells
→ Exocytosis - bulk transport out of cells

Endocytosis and exocytosis are processes that require energy and are forms of active transport

Endocytosis
Endocytosis
the bulk transport of liquids (pinocytosis) or solids (phagocytosis) into a cell, by the infolding of
the cell surface membrane to form vesicles containing the substance.

There are 2 forms of endocytosis:

• Phagocytosis:
→ Bulk intake of solid material by a cell.
→ Phagocytes are cells that specialise in phagocytosis.
→ The vacuoles formed are called phagocytic vacuoles.

• Pinocytosis:
→ Bulk intake of liquids by a cell.
→ If the vacuole (or vesicle) that is formed is extremely small then the process is called
micropinocytosis.
Exocytosis
Exocytosis
the bulk transport of liquids or solids out of a cell, by the fusion of vesicles containing the
substance with the cell surface membrane.

• The substances to be released (such as enzymes, hormones or cell wall building materials)
are packaged into secretory vesicles formed from the Golgi body or RER.
• These vesicles then travel to the cell surface membrane.
• Here they fuse with the cell membrane and release their contents outside of the cell.

Example
- the secretion of digestive enzymes from pancreatic cells.
- plant cells use exocytosis to get their cell wall building materials to the outside of the
cell surface membrane.
1. The rate of diffusion of a substance across a membrane increases as its concentration
gradient increases,
→ Diffusion shows a linear relationship, whereas facilitated diffusion has a curved
relationship with a maximum rate.
→ This is due to the rate being limited by the number of carrier proteins in the cell
surface membrane.

2. The rate of active transport also increases with concentration gradient,

→ It has a high rate even when there is no concentration difference across the
membrane.
→ Active transport stops if cellular respiration stops, since there is no energy.
Experiments
Investigating Diffusion
- Visking tubing is a non-living partially permeable membrane made from cellulose.
- Pores in this membrane are small enough to prevent the passage of large molecules (such as
starch and sucrose) but allow smaller molecules (such as glucose) to pass through by
diffusion.

Procedure
1. Filling a section of Visking tubing with a mixture of starch and glucose solutions

2. Suspending the tubing in a boiling tube of water for a set period time

3. Testing the water outside of the visking tubing at regular intervals for the presence of starch
and glucose to monitor whether diffusion of either substance out of the tubing has
occurred.

4. The results should indicate that glucose, but not starch, diffuses out of the tubing

This can be investigated more quantitatively by:

1. Estimating the concentration of glucose that has diffused into the water surrounding the
Visking tubing at each time interval .
(separate boiling tubes are set up for each time interval) using the semi-quantitative
Benedict’s test.)

2. Comparisons between the time intervals can be made with a set of colour standards (known
glucose concentrations) or a colorimeter to give a more quantitative set of results.

3. A graph could be drawn showing how the rate of diffusion changes with the concentration
gradient between the inside and outside of the tubing.
Surface Area to Volume Ratio (SA:V)
Principles of Surface Area to Volume Ratio
• Surface area
- refers to the total area of the organism that is exposed to the external environment.
• Volume
- refers to the total internal volume of the organism
(total amount of space inside the organism)
• As the surface area and volume of an organism increase
(and therefore the overall ‘size’ of the organism increases),
the surface area : volume ratio decreases.
- This is because volume increases much more rapidly than surface area as size
increases.

Calculating SA:V in different shapes

Investigating Surface Area
The effect of changing surface area to volume ratio on diffusion can be investigated by timing
the diffusion of ions through cubes of agar of different sizes.

Method
1. Coloured agar is made up and cut into cubes of the required dimensions .
(eg. [0.5cm x 0.5cm x 0.5cm], [1cm x 1cm x 1cm] and [2cm x 2cm x 2cm])

2. Purple agar can be created if it is made up with very dilute sodium hydroxide solution and
Universal Indicator.
Alternatively, the agar can be made up with Universal Indicator only.

3. The surface area, volume and surface area to volume ratio of these cubes is calculated and
recorded.

4. The cubes are then placed into boiling tubes containing a diffusion solution (such as dilute
hydrochloric acid).

5. The same volume of dilute hydrochloric acid should be carefully measured out into each
boiling tube.

6. The acid should have higher molarity than the sodium hydroxide so that its diffusion can
be monitored by a change in colour of the indicator in the agar blocks.

7. Measurements can be taken of either:

- The time taken for the acid to completely change the colour of the indicator in the
agar blocks
- The distance travelled into the block by the acid (shown by the change in colour of
the indicator) in a given time (eg. 5 minutes)
Analysis
1. If the time taken for the acid to completely change the colour of the indicator in the agar
blocks is recorded, these times can be converted to rates.

2. A graph could be drawn showing how the rate of diffusion (rate of colour change) changes
with the surface area : volume ratio of the agar cubes.
Investigating Osmosis
- This apparatus could be used to investigate how temperature or different concentrations of
solutions (differences in water potential) affect the rate of osmosis.

- As water moves into the concentrated sugar solution by osmosis, the level of water in the
narrow glass tube rises.

- The rate of this rise is a measure of the rate of osmosis.

Estimating Water Potential in Plant Tissue

• It is possible to investigate the effects of immersing plant tissue in solutions of different

water potentials and then use the results to estimate the water potential of the plant tissue
itself

• The most common osmosis practical of this kind involves cutting cylinders of potato and
placing them into solutions with a range of different water potentials
(usually sucrose solutions of increasing concentration – at least 5 different concentrations
are usually required)
Method
1. The required number of potato cylinders are cut .
(one for each of the solutions you are testing – or more than one per solution if you
require repeats)

2. They are all cut to the same length and, once blotted dry to remove any excess moisture,
their initial mass is measured and recorded before placing into the solutions.

3. They are left in the solutions for a set amount of time (eg. 30 minutes), usually in a water
bath (set at around 30oC).

4. They are then removed and dried to remove excess liquid.

5. The final length and mass of each potato cylinder is then measured and recorded.
Analysis
• The percentage change in mass for each potato cylinder is calculated

• The concentration of sucrose inside the potato cylinders can be found by:
1. Plotting a percentage change in mass against the concentration of sucrose solution
graph.
2. The point at which the line of best fit crosses the x-axis is the concentration of
sucrose inside the potato cylinders.
• If there is a potato cylinder that has neither increased nor decreased in mass, it means
there was no overall net movement of water into or out of the potato cells
- This is because the solution that this particular potato cylinder was in had the same
water potential as the solution found in the cytoplasm of the potato cells.
- There was no concentration gradient .
- Therefore no net movement of water into or out of the potato cells.
• A positive percentage change in mass indicates that the potato has gained water by osmosis
- The solution had a higher water potential than the potato.
- There is a net movement of water from the solution into the potato.
- This would make the potato cells turgid, as the water exerts turgor pressure (or
hydrostatic pressure) on the cell walls – the potatoes will feel hard.
• A negative percentage change in mass indicates that the potato gas lost water by osmosis
- The solution had a lower water potential than the potato.
- There is a net movement of water from the potato into the solution.
- This would make the potato flaccid and decreasing the mass of the potato cylinder –
the potato cylinders will feel floppy
- If looked at underneath the microscope, cells from this potato cylinder might be
plasmolysed.