IB Biology HL

Annie Lloyd
23/2/20

Osmosis In Onion Cells

Research Question:​ W​hat is the cytoplasm solute concentration of​ ​Allium cepa?​ ​?

Results
Table 1:​ Effects of Different Concentrations of Sodium Chloride Solution on the Recorded Number
of Plasmolysed Cells & Cells in Field of View
Concentr Number of Cells Counted (Cells) 土7
ations of
NaCl Trial 1 Trial 2 Trial 3 Average Range
(M) Plasmol of
ysed Plasm
% olysed
Plasmolysed Total Plasmolysed Total Plasmolysed Total %

0.00 0.00 60.00 0.00 43.00 0.00 65.00 0.00 0.00

0.25 5.00 59.00 3.00 57.00 2.00 48.00 5.97 4.31

0.50 11.00 42.00 20.00 60.00 11.00 25.00 34.51 17.81

1.00 83.00 112.00 76.00 100.00 127.00 146.00 79.03 12.88

1.50 103.00 118.00 98.00 121.00 104.00 115 86.24 9.44

2.00 138.00 147.00 109.00 112.00 121.00 123 96.53 4.7

The plasmolysed cells were identified to have an increased intensity of red colouration caused by a
higher concentration of pigment as water is diffused.The cell walls maintained their shape however
the cell contents shriveled up being observed as a dark red globule within the walls of the cell. In
comparison to this, cells that have not been plasmolysed are seen as having a lower concentration of
red pigment making the cells appear pink and the cell contents reaches all the way to the cell wall thus
they are filling are larger volume of space.

The above table communicates the collected data and the uncertainty of 土7 cells. When counting the
cells in the field of view and plasmolysed cells often fatigue occurs allowing the experimenter to
easily miscount cells. In addition to this it is hard to ensure that plasmolysed cells are not counted
twice thus there is slight uncertainty in the number of cells that have been identified as having been
plasmolysed. This is a limitation caused by human behaviour yet it does not have a great significance
to the recorded data. The uncertainty is relatively small compared to the number of cells being
counted thus not greatly affecting the accuracy of the data. Furthermore the uncertainty has a value of
7 as analysing the plasmolysed cells is subjunctive and can occasionally be difficult to determine
which cells have been through the process of osmosis and which cells have had structural damage
caused by the mechanical action of removing the onion layer. The leakage of pigment into the liquid
on the wet mount can often make it difficult to decipher cells but this has been taken into account. The
limitations of instruments also meant uncertainty occured in how much liquid was used per wet mount
as the pipette may have been slightly inaccurate however this error is so minimal. Furthermore the
 IB Biology HL
Annie Lloyd
23/2/20

microscopes resolution limitations limited the experimenter from noticing specific changes in a cell
structure making it hard to determine whether the cell was plasmolysed or not.

Discussion
Figure 1 illustrates the positive correlation between the concentration of sodium chloride and the
average number of plasmolysed cells expressed as a percentage of the total number of cells in the
microscope's field of view. The graph maintains an increase yet it is not consistent as shown by the
sudden rise between 0.25M and 0.5M where the percentage of plasmolysed cells changes by 28.54%.
This inconsistent growth continues as there is a growth of 44.52 % between the concentrations 0.5M
and 1M but this is to be expected as the concentration level changes by 0.5 M opposed to 0.25 M.
Between 1M and 2M consistent growth is noted with approximately a 10% growth every 0.5 M
increase in concentration. The continuous increase in this graph demonstrates how the sodium
chloride is a hypertonic solution compared to the cytoplasm solute of onion cells thus resulting in
water diffusing from the onion cells. The higher the concentration of sodium chloride the lower the
concentration of water thus more water is needed to balance the concentration levels of the two
substances. This results in more cells going through the process of osmosis to diffuse water out of the
cell which can be observed as an increase in the percentage of plasmolysed cells.
The error bars in the above graph do not overlap thus confirming the reliability and consistency of the
data. It is important to note that the errors for 0 M and 2 M are incorrect as <0% or >100% is not an
observable figure in this experiment.
 IB Biology HL
Annie Lloyd
23/2/20

Sample Calculations
Average %
Formula Worked Example

X = (11/42 + 20/60 + 11/25) * 100 ÷ 3

Range
Formula Worked Example

R = maximum value − minimum value R = (20/60 − 11/42) * 100

Evaluation
Strengths
A strength of this experiment was the use of technology to capture photographic evidence of the
number of cells; allowing the data to be checked multiple times and providing lasting qualitative data.
The experiment was consistent as different locations on the same wet mount were photographed and
analysed. This enabled averages to be found making the data more reliable. In addition it was
relatively easy to note the plasmolysed cells as the colour was noticeably darker and the cell
membrane had shrunk slightly, a benefit of using red onion cells opposed to dyed brown onion cells.
Furthermore this experiment yields itself to quantitative data which allows for inferential statistical
calculations to be made. This makes it easier to compare the data and demonstrates the validity of the
experiment by noting the average range of plasmolysed cells in the different concentrations being
relatively small. When the range between the three different trials is smaller it suggests that the results
are reliable.
The experiment is also relatively easy to repeat because the size of the onion layer used does not
affect the data thus valid repeats can be conducted and results obtained that concur with the
conclusion drawn from this experiment.
 IB Biology HL
Annie Lloyd
23/2/20

Weaknesses
This experiment was weak in that it relied heavily on the careful counting of cells where many data
measurement mistakes may have occurred but as discussed the uncertainty would not have a large
significance to the data.
Another weakness was the amount of time needed to extract the onion layers which was extremely
difficult and time consuming making the experiment rushed and causing damage to the cells. Often an
onion layer would fold over itself requiring more manipulation of the layer causing more damage.
This resulted in varying numbers of total cells as damaged cells were not included in the final
count.These ruptured cells caused pigment to leak into the mounting fluid making it slightly harder to
observe and count the plasmolysed cells.
Furthermore air bubbles still formed despite the careful laying off the slide cover using adhesive
forces. Although this does not greatly affect the collection of data it damaged some of the cells thus
osmosis could not be observed in these particular cells. This has a minimal significance on the data as
it was accounted for in the uncertainty.
It is important to also note that the onion cells and liquids may not have all been kept at the same
temperature having a slight impact on the results. Typically a higher temperature of substances
increases the rate of osmosis thus making it harder to compare the effect different concentrations have
on the amount of osmosis as the temperature factor was not controlled and the mounts were observed
after two minutes; not once all osmosis had been completed. Furthermore cell layers were removed
and left on the bench, causing oxidation of some layers thus they may have dried out, changing the
amount of water in individual cells and making it harder to observe the plasmolysed cells. This made
it difficult to attribute the changing rate of osmosis to the different concentrations of sodium chloride.
It is also hard to confirm if all onion cells in the wet mount were exposed to the sodium chloride
solution. As only a few drops were used some parts of the wet mount could have been studied where
the cells had little contact to the solution in comparison to other parts of the slide where they may
have been a larger volume of liquid affecting the amount of osmosis. This is a minimal problem as by
studying three different places on the slide then calculating an average it removes any possible error.

Improvements
To increase the reliability of this experiment three different pieces of onion layer could be studied per
concentration, reducing any errors and allowing for more substantial data to be collected creating
reliable averages. In addition to this the onion layer could be soaked in a beaker for longer than two
minutes to ensure that all onion cells are exposed to the solution making the osmosis process a lot
clearer and easier to study. If possible it is advised to extract a larger proportion of onion layer
enabling for more data to be explored as occasionally in the microscope's field of view the end of the
cell layer was photographed. This causes inconsistency in the total number of cells in the field of view
but this problem can be averted by using a larger layer of onion ensuring that there is ample amount of
cells to study and photograph without studying the same area.

In addition to this, to minimize membrane damage and reduce contact; the transparent inner layer of
the onion could be used which is easier to extract. It is necessary for this layer to be died with iodine
to make plasmolysis observable and would provide more data with fewer damaged cells.
 IB Biology HL
Annie Lloyd
23/2/20

To improve the validity of the experiment the onion layers could be stored in water to keep them
equally hydrated and maintain a consistent temperature to ensure that no external factors affect the
rate of osmosis thus providing more reliable data on how the concentrations of sodium chloride affect
the rate of osmosis.

Conclusion
The cytoplasm solute concentration of​ ​Allium cepa​ is estimated to be 0.65M as this is when 50% of
cells would show plasmolysis as shown in Figure 1. When only 50% of the cells show signs of
plasmolysis it can be assumed that it is the “equal point of incipient plasmolysis” (Cambridge
University Press 2008) where the solute concentration must be equal to the concentration of the cell’s
cytoplasm. The “equation of water relationships in plant cells is y​cell​ = y​s​ + y​p​; when no inward pressed
is caused by the cell wall (y​p​=0) the water potential of the cell equals the solute potential” (Yin Dip,
2005, University of Hong Kong). Plotting the percentage of plasmolysed cells against theq1e``2q a
concentrations of sodium chloride results in a line where the molarity of the solution can be
determined when 50 per cent of cells are plasmolysed. This evidence shows that the concentration of
the cytoplasm solute of ​Allium cepa​ is approximately 0.65M.

Extension
To better observe how different salt solutions affect the amount of osmosis that occurs potatoes could
be added to different salt solutions causing the potato to shrink and lose its turgor pressure. This will
provide more accurate data as the bigger difference in size and weight of the potato slice the higher
the concentration. The numerical data will be recorded through the use of scales and measuring
instruments that will have a smaller uncertainty than counting cells with the human eye. This data can
be used to calculate how different concentrations of sodium chloride affect the amount of osmosis that
occurs.
In addition other solutions with different concentrations could be used to see a stronger plasmolysis
with higher salt concentration. This will reflect the water potential of the onion cells concurring with
the conclusion made in this experiment.

An interesting comparison can also be made between how osmosis affects plant and animal cells
differently by placing red blood cells in solution of different osmolarities to demonstrate the effect of
osmosis and the resulting changes in cell volume. With the use of normal saline, observations can be
made as to how the different concentration levels affect blood cells differently.
 IB Biology HL
Annie Lloyd
23/2/20

Bibliography
Beck, William A. “Determining the Osmotic Value at Incipient Plasmolysis.” ​Transactions of the
American Microscopical Society,​ vol. 48, no. 2, 1929, pp. 204–208. ​JSTOR,​ accessed 24 Feb
2020.​www.jstor.org/stable/3222213

COAS Biology 1 Resource. (2008). ​Estimating the solute potential of cell sap by the incipient
plasmolysis method​. [online] [Accessed 23 Feb. 2020].

http://essentials.cambridge.org/media/COAS_B1_02_acts_pr5.pdf

Microbehunter.com. (2008). ​Observing Plasmolysis – Microbehunter Microscopy​. [online] [Accessed

Yip, Din. (2005). A reappraisal of the investigation on estimating the solute potential of plant cells.
School Science Review, accessed 22 Feb 2020
https://www.researchgate.net/publication/273257147_A_reappraisal_of_the_investigation_on_estimat
ing_the_solute_potential_of_plant_cells