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Anna Roman

AP Biology

Mrs. Schulte

Experiment #2

December 13-18th

The effect of different concentrations of solutions on the movement of water in and out of

various cells
Abstract:
The purpose of this experiment was to observe osmosis and diffusion in different
situations. We were able to fulfil the purpose by designing multiple experiments with real and
makeshift cells.
In procedure one we used dialysis tubing to create our own visible cells to understand
osmosis at at greater depth. We randomly chose solutions to put inside the tubing and solutions
the tubing would soak in. After preparing the solutions and cells we weighed them and let them
soak in the designated solution for 30 minutes. We used water in a water solution as our control
and tested NaCl in ovalbumin, glucose in NaCl, sucrose in ovalbumin, and glucose in sucrose.
After soaking, we removed the cells from their solution, dried off the excess liquid, and weighed
them. With the initial and final weight we were able to calculate the percent change and
determine if the solutions are hypotonic, hypertonic, or isotonic. Hypotonic solutions occur when
there is a higher concentration of solute inside the cell causing the water to move into the cell to
balance the concentrations. A hypertonic solution occurs when there is a higher solute
concentration outside the cell which causes water to move out of the cell and shrink. An isotonic
solution occurs when there is an equal solute concentration inside and outside the cell meaning
there is either no movement in or out of the cell or the movement equal each other. We
hypothesized that the cells would lose weight in a hypertonic solution due to movement of water
out of the cell, gain weight in a hypotonic solution due to the movement of water into the cell,
and remain the same when placed in an isotonic solution due to an equal net movement of water.
Based on the results we were able to determine that the water pair was an isotonic solution, the
ovalbumin and NaCl, NaCl and glucose, and ovalbumin and sucrose pairs were hypertonic
solutions and the sucrose and glucose pair was a hypotonic solution.
In the first part of procedure two we observed the movement of water in and out of a
freshwater plant after soaking in solutions with various solute concentration. Our hypothesis was
that we would see a change in the vacuole size and a separation of the cell wall and cell
membrane based on the type of solution the leaves were placed in. We looked at a leaf of an
Elodea plant under the microscope after it had been sitting in water to see what it looked like in
optimal conditions. This puts the plant in an isotonic solution and gave us a starting point to
observe organelle changes. We expected the vacuole to shrink and the cell wall and membrane to
separate when placed in a hypertonic solution ,and an increase in vacuole size as well as cell size
when placed in a hypotonic solution based on the movement of water into and out of the cell. We
let a few leaves soak in four different solutions: sucrose, glucose, ovalbumin, and NaCl, for ten
minutes. After soaking we looked at the leaves under a microscope again to observe the change
in the vacuole and cell wall/membrane. A change in these organelles is what we will use to
determine a change in turgor pressure in the plant cell. We were able to determine that sucrose
and glucose were hypotonic solutions and caused the vacuole to increase in size. The ovalbumin
and NaCl created a hypertonic solution causing water to leave the cell. We were able to tell
based on the decrease of the vacuole and separation of the cell wall and cell membrane which
happens because of the shrinking of the cell due to loss of water.
In the second part of procedure two we designed our own experiment of placing pieces of
sweet potatoes into color-coded solutions of unknown concentrations of sucrose. By weighing
the potatoes before and after soaking and using water as our control we were able to calculate the
percent change and determine the type of solution the sucrose created and make an educated
guess as to the concentrations of each solution. Based on the results we were able to determine
that solutions A, B, C and water created a hypotonic solution which means the solutions had a
high concentration of sucrose. Solution D created a hypertonic solution which means it had a low
concentration of sucrose. Based on the data we can assume the order from highest to lowest
sucrose concentration is A, B, C, D.

Materials and Equipment used:


Procedure 1:
↣ Distilled or tap water
↣ 1 M sucrose
↣ 1 M NaCl
↣ I M glucose
↣ 5% ovalbumin (egg white protein)
↣ 20 cm-long dialysis tubing
↣ Cups
↣ Balances
Procedure 2a:
↣ Elodea plant in freshwater
↣ Forceps
↣ 4 plastic cups
↣ 1 M sucrose
↣ 1 M NaCl
↣ 1 M glucose
↣ 5% ovalbumin (egg white protein)
↣ Microscope
↣ 5 microscope slides and cover slips
↣ Timer
↣ Light with stand
↣ 4 pipettes
Procedure 2b:
↣ Potatoes, sweet potatoes, or yams
↣ Cork borers or french fry cutter
↣ Balances
↣ Metric rules
↣ Cups
↣ Color-coded sucrose solutions of different, but unlabeled, concentrations prepared by
your teacher
↣ 250 mL glass beakers
↣ Paper towels

Procedure and methods:


Procedure 1:
Step 1
Choose up to four pairs of different solutions. One solution from each pair will be in the model
cell of dialysis tubing, and the other will be outside the cell in the cup. Your fifth model cell will
have water inside and outside; this is your control. Before starting, use your knowledge about
solute gradients to predict whether the water will diffuse into or out of the cell. Make sure you
label the cups to indicate what solution is inside the cell and inside the cup.
Step 2
Make dialysis tubing cells by tying a knot in one end of five pieces of dialysis tubing. Fill each
“cell” with 10 mL of the solution you chose for the inside, and knot the other end, leaving
enough space for water to diffuse into the cell.
Step 3
Weigh each cell, record the initial weight, and then place it into a cup filled with the second
solution for theat pair. Weigh the cell after 30 minutes and record the final weight.
Step 4
Calculate the percent change in weight using the following formula: (final - initial)/initial x 100.
Record your results.

Living cell membranes are selectively permeable and contain protein channels that permit the
passage of water and molecules. In some respects, the dialysis tubing you used is similar to a cell
membrane, and you can use it to explore osmosis in greater depth. Think about the questions that
came p as you worked through the investigation. Using the available materials, design an
investigation to answer one of your questions. Have your teacher check your design irst.
Remember to record your results, and be sure to use appropriate controls.
In this part of the lab the independent variable would be various solutions placed in and
out of the cells and the dependent variable is the osmosis that occurs when these solutions
are placed inside each other.

Procedure 3a:
Step 1
Start by looking at a single leaf blade from either ​Elodea​ (a water plant) or a leaf-like structure
from ​Mnium hornum​ (a moss) under the light microscope.
Step 2
Test one of the four solutions from Procedure 2 and find out if what you predicted is what
happens. Be sure to record all of our procedures, calculations, and observations.
In this part of the lab the independent variable is the solutions that the ​Elodea​ plant soaks
in and the dependent variable is the effect that the solutions had on the organelles and
turgor pressure of the plant cell.
Procedure 3b:
Design an experiment to identify the concentrations of the sucrose solutions and use the solutions
to determine the water potential of the plant tissues. (You might want to review the information
on water potential described in understanding water potential.)
In this part of the lab the independent variable is the various solutions the potatoes soaked
in and the dependent variable is once again the movement of water in and out of the cells.
Results:
Procedure 2
4 pairs
1) Water and water
2) Ovalbumin and NaCl
3) NaCl and glucose
4) Ovalbumin and sucrose
5) Sucrose and glucose

Weight change of dialysis tubing cells


Solution Solution inside Initial weight Weight after Percent
outside of cell of cell soaking change

H​2​O H​2​O 8.0g 8.0g 0%

Ovalbumin NaCl 5.9g 5.6g -5.08%

NaCl Glucose 8.2g 7.6g -7.32%

Ovalbumin Sucrose 8.3g 6.9g -16.87%


Sucrose Glucose 11.7g 12.6g 7.69%

We created makeshift cells out of dialysis tubing and picked solutions to put inside and outside
to create hypothermic, hyperthermic, and isothermic solutions with water as our control. The
sucrose and glucose combination created a hypothermic solution causing liquid to move into the
cell, the water combination created a perfect isotonic solution, and the other combinations
created hyperthermic solutions causing the liquid to go out of the cell. After weighing the cells
before and after letting them soak in the solution we were able to calculate the percent change
with the equation: (final-initial)/initial x 100.

Procedure 3
Observing osmosis in a water plant (​Elodea​)
Solution ​Elodea​ was placed in Results/observations

Sucrose Large increase in vacuole size, cells appear


swollen and no separation of the cell wall and
membrane

Glucose Small increase in vacuole but not as much as


the sucrose solution, and again now
separation of cell wall and membrane

Ovalbumin Large decrease vacuole size, separation of cell


wall, and the cell size appears to be shrunken

NaCl Small decrease in vacuole size, separation of


cell wall and cell membrane but not as much
as in the ovalbumin solution
We started this study by looking at a leaf of the ​elodea ​under a microscope after being in water
to see the size of the vacuole and relationship of the cell wall and cell membrane in its normal
state. We then placed leaves of the plant into different solutions to observe the water movement
and change in the structure of the cell. After soaking we looked at the leaves under the
microscope and found that sucrose caused the greatest increase in water movement into the cell
meaning it was a hypothermic solution, and ovalbumin caused the greatest increase in movement
of water out of the cell meaning it was a hyperthermic solution.

Potato weight and percent change after soaking in various concentrations of sucrose
solutions
Solutions A B C D H​2​O

Initial potato weight 7.1g 6.3g 5.4g 3.6g 4.1g

Potato weight post soak 7.63g 6.61g 5.54g 3.50g 4.52g

Percent change 7.46% 4.92% 2.59% -2.78% 10.42%


We placed sliced pieces of sweet potatoes into colored solutions of various concentrations of
sucrose to observe the osmosis that takes place in these different solutions. By weighing the
potatoes before and after they soaked in the solution we were able to determine whether the
solutions were hypothermic, or hypertonic. We found that all of the solutions were hypothermic
except solution D. Using this information we are able to determine the range of sucrose
concentration in each of the solutions

Analysis/conclusion:
This experiment fulfilled its purpose by determining the effect of different concentrations
of various solutions on varying cells. We were able to observe the osmosis and diffusion in plant
cells and larger makeshift cells.
In procedure 1 we were given four solutions that we were to use to create various
situations to observe the movement of water in and out of the cells. We used water in water as
our control (pair 1) and decided which solutions we were going to place in the cells and which
ones we were going to place outside the cells. We chose to put 5% ovalbumin inside 1 M
NaCl(pair 2), 1 M NaCl inside 1 M glucose(pair 3), 5% ovalbumin inside 1 M sucrose(pair 4),
and 1 M sucrose inside 1 M glucose(pair 5). Using dialysis tubing as a our “cell membrane” we
poured the correct solutions into labeled plastic cups and tied the dialysis tubing with the correct
solutions in it to make a cell. The dialysis tubing acted as a semipermeable membrane and
allowed water to pass through it but not the other larger molecules in the solutions. After
weighing each cell we recorded the data and let the cells soak in their dedicated solutions for 30
minutes. Not knowing the concentrations of the solutions we made a general hypothesis about
what would happen to the cells in different solutions. We assumed the cells would lose weight in
a hypertonic solution because the water would pass through the tubing to the outside of the cell
to balance the concentrations. The cells would gain weight in a hypotonic solutions because the
water would move into the cell to balance the concentrations; however, we did not hypothesize
that any of the cells would swell so much they would burst as they were not in the solutions for a
long enough time and the difference in the concentrations was not that great. We knew the water
pair would create an isotonic solution and the weight would remain the same due to no net
movement of water because of equal concentrations in and out of the cell. After the cells had
soaked for 30 minutes we took them out, dried off any excess liquid, and weighed them again.
The water pair remained the same at 8 grams before and after soaking meaning out hypothesis
was correct and it did create an isotonic solution. The ovalbumin inside the NaCl decreased from
5.9 grams to 5.6 grams meaning it created a hypertonic solution. The NaCl inside the glucose
decreased from 8.2 grams to 7.6 grams meaning it also created a hypertonic solution. The
ovalbumin inside of the sucrose decreased the most from 8.3 grams to 6.9 grams giving us
another hypertonic solution. The sucrose inside of the glucose was our only increase, and went
from 11.7 grams to 12.6 grams giving us our only hypotonic solution. With this data we were
then able to calculate the percent change using the equation (final-initial)/initialx100. Pair one’s
percent change was 0%, pair two’s was -5.08%, pair three’s was -7.32%, pair four’s was
-16.87%, and pair five’s was 7.69%. Based on this data our hypothesis was validated and we
were able to determine which solutions had higher concentrations than its pair. We were also
able to observe osmosis on a visible scale.
In procedure 2 part a we used an ​Elodea​ plant to observe osmosis in a real life situation
and determine the effects changes to turgor pressure have on plant cells. We hypothesized that
different types of solutions would have different effects on organelles that play an important role
in water retention for the plant. We estimated that hypertonic solutions would cause the cell size
to shrink along with the vacuole size and the cell wall to separate from the cell membrane.
Hypotonic solutions would cause an increase in vacuole size as well as cell size. Our isotonic
solution would be the control or the ​elodea​ plant after soaking in its optimal conditions of
freshwater. We looked at a few of the plant leaves after soaking in freshwater under the
microscope to get a basis of what the plant looks like under normal circumstances to use as a
control for looking at the plant under unusual circumstances. After looking at the plant under the
microscope we then put a few leaves into cups of 1 M sucrose, 1 M glucose, 5% ovalbumin, and
1 M NaCl and let them soak for 10 minutes. After soaking, we placed a leaf on a microscope
slide and looked at it under a microscope to observe the changes in the cell compared to what we
saw the first time and recorded the data. We found the sucrose and glucose created a hypotonic
solution causing the water to move into the cell and the vacuole to increase and size and the cell
to appear swollen. The sucrose increased the most compared to the glucose. The ovalbumin and
NaCl creates hypertonic solutions and caused the water to leave the cell, the vacuole size to
shrink, and the cell membrane begin to come away from the cell wall. The ovalbumin has the
greatest change in cell size compared to the NaCl. Using this data we were able to determine the
effects the different solutions and change in turgor pressure have on a plant cell in real life.
In procedure 2 part b we were able to create our own experiment using sweet potatoes and four
solutions of sucrose with unknown concentrations with water as our control. We weighed the
potato slices before placing them in the sucrose solutions and the water and then let them soak
for 30 minutes. With a hypothesis similar to before we assumed the potato would lose weight in
a hypertonic solution, gain weight in a hypotonic solution and remain the same in an isotonic
solution. After soaking we weighed the potatoes and recorded the data. We found that the
potatoes in solutions A, B, C, and water increased and solution D decreased. Solution A went
from 7.1 grams to 7.63 grams, B went from 6.3 grams to 6.61 grams, C went from 5.4 grams to
5.54 grams, and the water went from 4.1 grams to 4.51 grams. These solutions created a
hypertonic solution and caused the water to move out of the potatoes. Solution D went from 3.6
grams to 3.5 grams which created a hypotonic solution. The water was not an isotonic solution
because sweet potatoes have a higher sugar concentration water; therefore causing the water to
move into the potato to balance the concentrations. Using this data we were able to calculate the
percent change using the same formula as from procedure one. Solution A changed 7.46%,
solution B changed 4.92%, solution C changed 2.59%, solution D changed -2.78%, and the water
changed 10.42%. We were also able to determine the order from highest to lowest of sucrose
solution as A, B, C, D.
A few sources
Background Research and Bibliography:
Cells must have materials through membranes and throughout cytoplasm in order to
maintain homeostasis. The movement is regulated because cellular membranes, including the
plasma and organelle membranes, are selectively permeable. Membranes are phospholipid
bilayers containing embedded proteins; the phospholipid fatty acids limit the movement of water
because of their hydrophobic characteristics.
The cellular environment is aqueous, meaning that the solutes 9e.g., salts, organic
molecules) dissolve in water, which is the solvent. Water may pass slowly through the membrane
by osmosis or through specialized protein channels called aquaporins. Aquaporins allow the
water to move more quickly than it would through osmosis. Most other substances, such as ions,
move through protein channels, while larger molecules, including carbohydrates, move through
transport proteins.
The simplest form of movement is diffusion, in which solutes move from an area of high
concentration to an area of low concentration; diffusion is directly related to molecular kinetic
energy. Diffusion does not require energy input by cells. The movement of a solute from an area
of low concentration to an area of high concentration requires energy input in the form of ATP
and protein carriers called pumps.
Water moves through membranes by diffusion; the movement of water through
membranes is called osmosis. Like solutes, water moves down its concentration gradient. Water
moves from areas of high potential (high free water concentration) and low solute concentration
to areas of low potential (low free water concentration) and high solute concentration. Solutes
decrease the concentration of free water, since water molecules surrounded the solute molecules.
The terms​ hypertonic, hypotonic​, and​ isotonic​ are used to describe solutions separated by
selectively permeable membranes. A hypertonic solution has a higher solute concentration and a
lower water potential as compared to the other solution; therefore, water will move into the
hypertonic solution through the membrane by osmosis. A hypotonic solution has a lower solute
concentration and a higher water potential than the solution on the other side of membrane; water
will move down its concentration gradient into the other solution. Isotonic solutions have equal
water potentials.
In nonwalled cells, such as animal cells, the movement of water into and out of a cell is
affected by the relative solute concentration of either side of the plasma membrane. As water
moves out of the cell, the cell shrinks; of water moves into the cell, it swells and may eventually
burst. In walled cells, including fungal and plant cells, osmosis is affected not only by the solute
concentration, but also by the resistance to water movement in the cell by the cell wall. This
resistance is called turgor pressure. The presence of a cell wall prevents the cells from bursting
as water enters; however, pressure builds up inside the cell and affects the rate of osmosis.
Water movement in plants is important in water transport from the roots into the shoots
and leaves. You likely will explore this specialized movement called transpiration in another lab
investigation.

Understanding water potential:


Water potential predicts which way water diffuses through plant tissues and is
abbreviated by the Greek letter psi (𝜓). Water potential is the free energy per mole of water and
is calculated from two major components: (1) the solute potential (𝜓​s​). Which is dependent on
solute concentration, and (2) the pressure potential (𝜓​p​), which results from the exertion of
pressure-either positive or negative (tension)- on a solution. The solute potential is also called the
osmotic potential.
𝜓=𝜓​p​+𝜓​s
Water potential=Pressure Potential+Solute Potential
Water moves from an area of higher water potential or higher free energy to an area of
lower water potential or lower free energy. Water potential measures the tendency of water to
diffuse from one compartment to another compartment.
The water potential of pure water in an open beaker is zero (𝜓=0) because both the solute
and pressure potentials are zero (𝜓​s​=0;𝜓​p​=0). An increase in positive pressure raises the pressure
potential and the water potential. The addition of solute to the water lowers the solute potential
and therefore decreases the water potential. This means that a solution at atmospheric pressure
had a negative water potential due to the solute.
The solute potential (𝜓​s​)= -iCRT, where i is the ionization constant, C is the molar
concentration, R is the pressure constant (R=0.0831 liter bars/mole-K), and t is the temperature
in K (273+ degrees celcius).
A 0.15 m solution of sucrose at atmospheric pressure (𝜓​p​=0) and 25 degrees celsius has
an osmotic potential of -3.7 bars and a water potential of -3.7 bars. A bar is a metric measure of
pressure and is the same as 1 atmosphere at sea level. A 0.15 M NaCl solution contains 2 ions,
Na​+​ and Cl​-​; therefore i=2 and the water potential= -7.4 bars.
When a cell’s cytoplasm is separated from pure water by a selectively permeable
membrane, water moves from the surrounding area, where the water potential is higher (𝜓=0),
into the cell, where water potential is lower because of solutes in the cytoplasm (𝜓 is negative). It
is assumed that the solute is not diffusing. The movement of water into the cell causes the cell to
swell, and the cell membrane pushed against the cell wall to produce an increase in pressure.
This pressure, which counteracts the diffusion of water into the cell, is called turgor pressure.
Over time, enough positive turgor pressure builds up to oppose the more negative solute
potential of the cell. Eventually, the water potential of the cell equals the water potential of the
pure water outside the cell (𝜓 of cell=𝜓 of pure water=0). At this point, a dynamic equilibrium is
reached and net water movement ceases.
If solute is added to the water surrounding the plant cell, the water potential of the
solution surrounding the cell decreases. If enough solute is added, the water potential outside the
cell is equal to the water potential inside the cell, and there will be no net movement of water.
However, the solute concentration inside and outside the cell are not equal, because the water
potential inside the cell results from the combination of both the turgor pressure (𝜓​p​) and the
solute pressure (𝜓​s​).
If more solute is added to the water surrounding the cell, water will leave the cell, moving
from an area of higher water potential to an area of lower water potential. The water loss causes
the cell to lose turgor. A continued loss of water will cause cell membrane to shrink away from
the cell wall, and the cell will plasmolyze.

Works Cited:

Tutor, S. (2017, April 25). What Happens to Plant & Animal Cells When Placed in Hypertonic,

Hypotonic & Isotonic Environments? Retrieved January 03, 2018, from

https://sciencing.com/happens-hypertonic-hypotonic-isotonic-environments-8624599.html