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Background Information
Osmosis is defined as the movement of solvent molecules across a selectively permeable
membrane. In osmosis, water moves into a region of higher solute concentration or down its
concentration gradient since, water can diffuse through a membrane. Osmosis is also a passive
process that does not use energy or ATP to pass through a cell membrane and therefore makes it
Tonicity describes the ability of a solution to impact the shape of a cell by influencing the
cells internal water volume. Tonicity is influenced by solutes that are unable to cross the cell
membrane and there are three types of tonicity solutions that are used to make comparisons to a
cell: Isotonic solutions, Hypertonic solutions and Hypotonic solutions (Hoehn 71). In Isotonic
solutions, human blood cells retain their normal size due to the fact that solute and water
concentrations are the same both inside and outside the cell and there is no net osmosis taking
place. In Hypertonic solutions, cells lose water by osmosis and shrink or shrivel because there is
a higher concentration of solutes on the inside of the cell. In Hypotonic solutions, cells gain
water by osmosis and swell and even burst or lyse because there is a lower concentration of
solutes on the inside of the cell (Marieb 59) (Science: Osmosis BBC).
This experiment will examine the impact of solutions with different tonicities on red
blood cells. The independent variable will be the red blood cells and the dependent variable will
be the varying solutions that the red blood cells are placed in. Three solutions will be used are
the dependent variable, 0.9% NaCl, 10%NaCl and DI H2O. The control variable will be the
unaltered blood. Diffusion will be observed and the results will be examined under a microscope
Purpose
The purpose of this experiment is to determine how red blood cells react when suspended in
Hypothesis
The red blood cells should shrink in the 10%NaCl, swell or lyse in the DI H2O and remain the
Procedure
Equipment Used
0.9% NaCl Wooden Applicator Stick 4 Clean Cover Slides
10%NaCl Disposable mat Cover slips
DI H2O Disposable Gloves Microscope
Sheep’s Blood Eye Wear Wax Pencil
Collection of Data
Place disposable matt or napkin on your table as well as four slides and cover-slips. A
Sample of sheep’s blood will be provided from the instructor. Number each slide with a wax
pencil to easily identify. Place a drop of blood on a slide and number it Slide #1.
With the wooden applicator stick, dip the end into the blood and transfer small amounts
Slide #2
1. Combine 1 drop of 0.9% NaCl with the blood already on the slide
2. Add a cover-slip on top and use the filter paper to absorb excess liquid (if necessary).
3. Examine the cells under the microscope at 40X, then 100X and then high power (400X).
Slide #3
1. Place 1 drop of 10% NaCl solution onto the slide with the drop of blood
2. Add a cover-slip and use the filter paper to absorb excess liquid (if necessary).
3. Examine the cells under the microscope at 40X, then 100X and then high power (400X).
Slide #4
1. Place 1 drop of DI H2O onto the slide with the drop of blood (use the filter paper to absorb
excess liquid).
2. Add a cover-slip and use the filter paper to absorb excess liquid (if necessary).
3. Examine the cells under the microscope at 40X, then 100X and then high power (400X).
Choose which osmotic process is being observed in each of the three experiments and label the
When finished with the experiment, place slides and cover slips into the beaker containing
bleach. Place all disposable items in the Solid Biohazard Waste Container. (Hill)
Results
Table
Description of Data
Slide 1 is the control or original sample. It is expected to be in an isotonic solution and will be
used as a baseline to compare with the dependent variable. Slide 2 contained Blood in 0.9%
NaCl Solution. This would make it an Isotonic solution and the red blood cells showed no
significant change. Slide 3 contained Blood in 10%NaCl. This would make the solution
Hypertonic and the cells appeared to shrivel. Slide 4 contained DI H2O. This would make the
solution Hypotonic and the cells appeared larger than the control.
Discussion
Support of Hypothesis
The results of the experiment matched the initial prediction. The hypothesis stated that the red
blood cells would shrink in the 10%NaCl, swell or lyse in the DI H2O and remain the same or
have no large change in 0.9%NaCl. Since the hypothesis prediction reflected the results of the
the effect of solutions of varying tonicities on red blood cells. By examining the changes in
shape that red blood cells undergo, insight can be gained into diffusion and osmosis.
The control, which was slide 1, had deep red cells in color because no additional solvent
was added to the solution. The cells of slide 1 appeared normal round and as the control, set the
standard size that we that we would compare the other samples too.
In slide 2, the cells were similar in size to slide 1 and also round. They did however lose a
very small amount of their dark red color, when compared to the other samples, since a drop of
0.9 NaCl was added to the blood. As a result, it was determined that this was an Isotonic
solution.
Slide 3 had a very significant change from the previous sample. Slide 3 contained
10%NaCl and blood. The cells appeared to be smaller than the previous slides and lost their
round shape to look more oval in appearance. The cell also lost a significant amount of their red
appearance and appeared clear when viewed through a microscope. As a result, this sample was
Slide 4 contained DI H2O and blood. In slide 4, the cells were larger and looked rounder
than all the other samples that we observed. In terms of color, the cells also looked similar in
solution.
The results of the experiment matched the expectations that were initially stated in the
hypothesis. The color changes that occurred in the blood cells were not a part of the hypothesis
but are noteworthy. The cells of Slide 3 three appeared to lose their color in the presence of 10%
NaCl. More research could be done to done determine why the cells on slide three lost their
color.
Clinical Application
This experiment provided information that can be used in the future in real life applications.
Medical professionals can often examine cells shapes to make an accurate diagnosis of a patient.
For example, if a medical professional believes that a patient is dehydrated he or she could
examine the blood and the shapes of blood cells to confirm his diagnosis. If the patient was
patient’s ion or salt levels were too high, you would also expect the blood cells to be shriveled. If
a patient’s ion levels were too low however, you would expect the patient’s blood cells to swell
or in critical conditions even lyse. It would be favorable for a medical professional to examine
tonicity when making a diagnosis because the process can be performed easily and affordably.
References
Hill, Heather. Investigating Diffusion Through Living Membranes. Clarkston: Georgia Perimeter
Hoehn, Katja. "Cells: The Living Units." Human Anatomy & Physiology Masteringa & P With
Etext. By Elaine N. Marieb. 9th ed. N.p.: Pearson College Div, 2015. 61-81. Print.
Marieb, Elaine Nicpon, Susan J. Mitchell, and Peter Z. Zao. "The Cell: Transport Mechanisms
and Cell Permeability." Human Anatomy & Physiology Laboratory Manual. 10th ed. San