Laboratory Practice 2

“Water transport across cell membranes”

GROUP MEMBERS
SUBJECT Biology SL DP
YEAR/GRADE IV DP
Biol. Gen. Biotechnol.
TEACHER
Andres Agurto

1. Introduction:
Water transport across cell membranes is a fundamental biological process crucial for
maintaining cell homeostasis. Understanding how cells respond to different tonicity
environments is essential in both plant and animal biology. In this laboratory experiment, we
will investigate water transport in plant cells using a plant sample (such as Elodea or
Tradescantia leaf) and in animal cells using blood extracted from students. By exposing
these cells to isotonic, hypotonic, and hypertonic solutions, we can observe the effects of
osmosis on cell shape, size, and overall cellular health.
The experiment involves examining the behavior of plant and animal cells when exposed to
isotonic, hypotonic, and hypertonic solutions. For the plant cell experiment, we will observe
the changes in cell structure and morphology as the cells are subjected to different tonicity
conditions. This will provide insights into how plant cells adapt to varying osmotic pressures.
Similarly, in the animal cell experiment, we will investigate the impact of osmotic imbalances
on red blood cells derived from student blood samples. By carefully analyzing the alterations
in cell shape and size, we can gain a better understanding of how animal cells respond to
different tonicity environments. Through these experiments, we aim to explore the
mechanisms of water transport across cell membranes and the effects of osmosis on plant
and animal cells.
Design
2.1. Objectives:

• Objective 1: To observe and compare the changes in plant cell morphology when
exposed to isotonic, hypotonic, and hypertonic solutions. This objective aims to
investigate the effects of osmosis on plant cells and understand how they adapt
to different tonicity environments.
• Objective 2: To examine and analyze the alterations in red blood cell shape and
size when subjected to isotonic, hypotonic, and hypertonic solutions. This
objective focuses on understanding the response of animal cells to osmotic
imbalances and studying the impact of tonicity on red blood cells.
 • Objective 3: To gain a comprehensive understanding of water transport across
cell membranes through the processes of osmosis and osmoregulation. By
observing the behavior of plant and animal cells under different tonicity
conditions, this objective aims to elucidate the mechanisms by which cells
maintain homeostasis and regulate water balance.
• Overall, the objectives of this laboratory practice are to explore the effects of
osmosis on plant and animal cells, investigate the changes in cell morphology
and behavior under isotonic, hypotonic, and hypertonic conditions, and deepen
our understanding of water transport across cell membranes.

1.2 Research question:

1.3 Hypothesis formulation:

2. Methods
2.1 Materials

Biological material
Laboratory materials per table Reagents (Provided by the
student)

• Plant sample
• Isotonic solution (e.g., (Elodea)
0.9% saline solution) • Microscope slides
• Microscope and coverslips
• Hypotonic solution
• Dropper or pipette • Glass marker
(e.g., distilled water)
• Microscope slide holder or a • Sterile puncture
• Hypertonic solution
petri dish lancet
(e.g., 1.5% saline
• Microscope • Nitrile Gloves
solution)
 2.2 Procedure:
Preparation:
a. Label each microscope slide with the sample type and solution concentration
(isotonic, hypotonic, hypertonic).
b. Place the microscope slide holder or a petri dish on your work area.
c. Fill three separate containers with distilled water, isotonic solution, and hypertonic
solution.
Plant Cell Experiment:
a. Take a fresh leaf sample from the plant, ensuring it is healthy and intact.
b. Place the leaf sample on a microscope slide and gently press a coverslip on top to
flatten it.
c. Add a drop of distilled water to the slide and gently mix it with the leaf.
d. Observe the cells under a microscope using low magnification to locate an area with
intact cells.
e. Switch to higher magnification and focus on a single plant cell.
f. Record your observations of the cell, noting its size, shape, and any visible structures
(e.g., vacuoles, cell wall).
g. Remove the coverslip and carefully add a drop of hypotonic solution to the slide.
h. Replace the coverslip and gently press down to ensure the solution spreads evenly.
i. Observe the cell under the microscope, focusing on any changes in cell shape or
size.
j. Record your observations of the cell in the hypotonic solution.
k. Repeat steps g-j for the isotonic and hypertonic solutions, using fresh slides and
labeled containers for each solution.
l. Compare and document the changes observed in the cells under different solutions.
Animal Cell Experiment:
a. Obtain a blood sample from the students using appropriate methods and
precautions.
b. Place a drop of blood on a microscope slide, spreading it out evenly.
c. Allow the blood to dry completely on the slide or fix it using suitable staining
techniques (if desired).
d. Add a drop of distilled water to the slide, covering the blood sample.
e. Gently place a coverslip on top of the water drop, ensuring no air bubbles are
trapped.
f. Observe the slide under a microscope using low magnification to locate an area with
intact red blood cells.
g. Switch to higher magnification and focus on a single red blood cell.
h. Record your observations of the cell, noting its shape, size, and any visible structures
(e.g., cell membrane, cytoplasm).
i. Remove the coverslip and carefully add a drop of hypotonic solution to the slide.
j. Replace the coverslip and gently press down to ensure the solution spreads evenly.
 k. Observe the red blood cell under the microscope, focusing on any changes in cell
shape or size.
l. Record your observations of the red blood cell in the hypotonic solution.
m. Repeat steps i-l for the isotonic and hypertonic solutions, using fresh slides and
labeled containers for each solution.
n. Compare and document the changes observed in the red blood cells under different
solutions.

3 Questionnaire

• Question 1: What were the observable changes in plant cell morphology when
exposed to isotonic, hypotonic, and hypertonic solutions? Describe any alterations
in cell shape, size, or visible structures.
• Question 2: How did the red blood cells extracted from student blood samples
respond to isotonic, hypotonic, and hypertonic solutions? Explain any changes
observed in terms of cell shape, size, or overall cellular health.
• Question 3: Which solution (isotonic, hypotonic, or hypertonic) caused the most
significant alterations in plant cell morphology and why? Provide a possible
explanation based on the principles of osmosis.
• Question 4: Did you observe any notable differences between the responses of plant
cells and red blood cells to the isotonic, hypotonic, and hypertonic solutions? Discuss
any similarities or dissimilarities in their behavior.
• Question 5: Based on your observations and understanding of water transport across
cell membranes, explain the importance of osmosis and osmoregulation for the
maintenance of cellular homeostasis in both plant and animal cells.

4 References
• Campbell, N. A., Reece, J. B., Urry, L. A., et al. (2008). Osmosis and water potential
in plants. Biology (8th ed.).
• Verkman, A. S. (1996). Osmoregulation in animal cells. American Journal of
Physiology-Cell Physiology, 270(3), C435-C455.
doi:10.1152/ajpcell.1996.270.3.C435
• Taiz, L., & Zeiger, E. (2010). Plant cell turgor and water stress. In Plant Physiology
(5th ed.)..
LABORATORY REPORT OUTLINE
1. Introduction
2. Design: objectives, research question, formulation of hypotheses, variables.
3. Methodology: materials, procedure
4. Results: includes all the observations made during the practice.
5. Discussion
6. Conclusions
7. Questionnaire
8. References