Cellular Transport Systems

Bautista, J.E., Calicdan, J.L.D., Carena, J. V., Casipit, L.A.B., & Damian, S. J. B.

Department of Biology

College of Science

University of the Philippines- Baguio City

Abstract

The cell membrane controls the movement of molecules in the cell and is essential for the

survival of life. This experiment aims to observe the structure of the plasma membrane, its

selective permeability in terms of the factors that affects the said feature. It also aims to

determine the factors that affect diffusion and, the behavior of plant and animal cells during

osmosis, particularly on their exposure to hypotonic, hypertonic and isotonic solutions. The

structure of the plasma membrane was observed on the oil globules formed on the oil-water

mixture. The selective permeability was observed with the use of yeast cell with formalin,

applied with heat, and a control. Diffusion was observed with the use of starch and glucose

solution in a collodion tube and was immersed in an iodine solution. For osmosis, Boat of

Mosses (Tradescantia spathacea) was used for plant cells and human red blood cells for animal

cells were used. It was concluded that the cell membrane is highly selective with the passing

materials. Heat increases the permeability and while formalin limits the movement of materials.

The cell shape and size is also affected by osmosis, specifically the concentration of the

extracellular environment. The diffusion of materials is affected by the molecule’s size.

INTRODUCTION

All living things have cells. Even if it is the smallest unit of life, its processes and

structures is vital for the survival of primitive life forms up to the most complex of organisms.

One of the most important features of the cell essential for survival is the plasma

membrane. The structure of the plasma membrane was first presented through the model

proposed by Jonathan Singer and Garth Nicholson. The Fluid-Mosaic Model illustrates the

components of the plasma membrane wherein protein molecules attached with carbohydrate

molecules are embedded in the lipid bilayer (Singer & Nicolson, 1972). These proteins can move

around and “float” among the semi-fluid lipids. Because the cell membrane is made up of many

different kinds of molecules which resembles a “mosaic”, scientists describes the cell membrane

as a “fluid-mosaic” structure (Miller & Levine, 2010).

The cell membrane is a phospholipid bilayer of hydrophilic heads of glycerol-phosphate.

Since phosphate groups give end polarity and are attracted to the polar water molecules, the

hydrophilic heads at the outer and inner parts of the bilayer points away and towards the cell,

respectively. On the other hand, non-polar hydrophobic fatty acid tails points toward each other

to form an oily layer inside the membrane that keeps the water out. Phospholipids automatically

arrange themselves in the way that hydrophobic tails are pointing on each other while

hydrophilic heads are facing the areas inside and outside of the cell when they are scattered in

water (Crash Course, 2012).

Due to the complex structure of the cell membrane, it enables the cell to be selectively

permeable allowing various types of molecules to be transported in two different mechanisms

either assisted or unassisted.

 The plasma membrane, also called the cell membrane, is a membrane found in all cells

that separates the interior of the cell from the outside environment serving as a protection against

foreign invaders such as bacterial cells. It also regulates the transport of materials entering and

exiting the cell which helps in sustaining cellular equilibrium (Green, n.d.). The mechanisms that

molecules are transported across the membrane are highly dependent on various factors such as

molecular size, polarity, and molecular charge. Usually, small, non-polar, and uncharged

molecules can easily pass through the membrane. The way these molecules are transported is

through unassisted membrane transport mechanism.

On the other hand, molecules that are large, polar, and charged are transported with the

use of channel proteins and carrier/transport proteins which utilizes energy. This mechanism is

called assisted membrane transport.

The different transport mechanisms illustrate the selective permeability of the cell and its

action to regulate its water and solute content, nutrient absorption, and regulation of cellular

materials.

The objectives of this experiment are to observe the structure of the plasma membrane,

its selective permeability in terms of the factors that affects the said feature. It also aims to

determine the factors that affect diffusion and, the behavior of plant and animal cells during

osmosis, particularly on their exposure to hypotonic, hypertonic and isotonic solutions.

MATERIALS AND METHODS

The group divided the experiment into four parts to determine the structure of the plasma

membrane, its selective permeability and the factors that affects it, the factors that affect

diffusion, and the behavior of the plant and animal cells in diffusion.

A. Structure of the plasma membrane

In a test tube, five (5) ml of oil and five (5) ml of tap water were mixed vigorously. It was

then placed it in a test tube rack to be observed after a three (3) minutes. After observation, the

test tube was once again mix without allowing the oil and water layers to settle. Several drops of

the oil and water mixture were placed in clean glass slide. The interphase between the oil and

water is then examined under Low Power Objective (LPO) for recording of data and

interpretation.

B. Selective action of cell membranes

The group dropped one (1) ml of aqueous yeast suspension in each of the three (3) test

tubes labeled 1, 2, and 3. Three drops of Congo red solution were mixed to each test tube. For

the experimental groups, four drops of 40% formalin were added to test tube 1, while for test

tube 2, it was placed in a hot water bath until it is near boiling point. The test tube 3 serves as the

control. Next, two drops of each suspension were placed on glass slides and placed under the

microscope for observation and, recording of data and interpretation.

C. Diffusion

With the prepared collodion tubes, the diffusion of the iodine solution is observed. The

prepared collodion tubes contain five (5) ml of 80% starch solution (collodion tube 1) and 5 ml

of 80% glucose solution. These are then immersed in 80mL water with 20mL 2% iodine
solution. The changes in color if the inside and outside of the collodion tube was then observed

and recorded for data interpretation.

D. Osmosis

I. Plant cells

A water mount of a Boat of Moses (Tradescantia spathacea) leaf was prepared,

specifically the violet portions, using distilled water. Then, the size of the cell was examined

under the microscope under High Power Objective (HPO). After viewing under the microscope,

the distilled water was then drained. Next, the group placed a drop of 0.5% NaCl solution on the

side of the c and allowed the solution to penetrate the specimen. The changes on the size of the

cell were observed and recorded. Afterwards, a new water mount was prepared and dropped with

10% NaCl and was also observed under the microscope for recording and interpretation.

II. Animal Cell (Human red blood cell, RBC)

A few drops of blood was extracted and placed on two clean glass slides. This is then

dropped with 0.9% NaCl solution (first slide) and distilled water (second slide). The two slides

are then examined specifically the size and shape of the red blood cells for recording of data and

interpretation. Afterwards, the second slide with distilled water and blood was dropped with

10%NaCl solution. This is then also examined under the microscope.

RESULTS AND DISCUSSIONS

This further elaborates the outcomes which were gathered while doing the laboratory

activity. Additionally, this discusses the questions presented in the different parts of the

laboratory activity in order to support the ideas regarding it.

A. Structure of the Cell Membrane

After the water and oil mixture was vigorously shaken, the oil globules did not mix with

the water molecules and they are seen in large circles under the microscope. They are scattered

around in large and uneven circles which are immiscible to the water molecules.

Figure 1. Water and oil mixture under the microscope

The oil globules in figure 1 can be a representation of the behavior of the plasma

membrane that has been essential to life. The phospholipids that comprise the plasma membrane

are amphipatic molecules having a hydrophilic polar heads and hydrophobic non-polar tails.

Polar molecules, like water, are strongly attracted to the hydrophilic head. However water cannot

easily pass through the membrane because of the hydrophobic non-polar tail of the phospholipid.

The non-polar part is composed of a saturated and unsaturated chain of fatty acid. Fats and water

cannot mix and since fats are nonpolar and water is a polar molecule. Thus, this property of the

cell membrane makes the amoeba acquires a barrier to protect and make it intact from the aquatic

medium where it is placed.

B. Selective action of the cell membranes

The cell membrane regulates the flow of materials in and out of the cell. It possesses

certain selectivity with which it maintains a chemical balance between the cell and its

environment. The lipid bilayer provides the basic fluid structure of the membrane and serves as

relatively impermeable barrier to the passage of most water soluble molecules (Alberts,

et.al.,2009).

Congo Red

Test Tube 1 Test Tube 2 Test Tube 3

40% Formalin Heat No Treatment

Yeast

Solution

Table 1. Three test tubes under different conditions

The results further illustrate the selective action of the cell membrane. In the three test

tubes which were subjected for experimentation, the cells were most stained at the test tube

subjected to heat compared to the test tube containing 40% formalin and the controlled test tube.

The test tube containing formalin did not accept the dye.
 The movement of materials in and out of the cell is affected by various factors. High

temperature allows materials to move in and out of the cell easily, illustrated by the reaction of

the dye with the yeast cells when heated. Increasing the temperature causes the phospholipids to

move, making the bilayer partially permeable. When heat is introduced in the treatment, proteins

in the membrane are denatured resulting to membrane being disrupted, too. Thus this allowed the

dye to enter the yeast cells in the heated treatment. The test tube exposed to heat has more dyed

yeast cells than the test tube with formalin and the one applied with not applied with any

treatment.

The yeast cells were not dyed when exposed to the formalin solution. This is due to the

process of fixation. According to Thermofisher Scientific (n.d.), formalin bonds adjacent

macromolecules, such as the integral proteins in the plasma membrane that facilitates the

movement of molecules such as water. This process is known as crosslinking. In this process, the

cell membrane’s permeability is high limiting the amount of molecules moving in and out of the

cell. Thus, the cell treated with formalin was not stained.

C. Diffusion

In this setup, the starch inside the collodion tube was stained by the iodine from the

iodine solution in the beaker. The iodine solution penetrated the collodion tube and there is a

color change from white to violet-purple because the iodine can stain polysaccharides. Starch is a

polymer of glucose which is composed of an amylopectin and amylose chain. Amylopectin is

known to react with iodine.

 Figure 2. Staining of five (5) ml of 80% starch solution in the collodion tube 1

The permeability of the collodion tube allows the smaller particles of iodine molecules to

pierce through while the starch molecules remained inside due to its relatively bigger size. The

smaller size of the iodine allows it to move freely through the membrane while the more

complex starch molecules cannot pass through it.

Figure 3. Staining of glucose in the collodion

 In this setup, the glucose inside the collodion tube was not stained by the iodine from

iodine solution in the beaker. The iodine solution penetrated the collodion tube but there were no

traces of any stain from the glucose inside the tube. It only acquired a brownish color from the

iodine which entered the collodion tube but the glucose remained as is. Even though starch and

glucose are both carbohydrates, iodine will not stain the glucose because it is just a

monosaccharide and does not contain amylopectin which reacts with iodine on the starch

solution.

The permeability of the collodion tube allows the smaller particles of iodine molecules to

pierce through while the glucose molecules remained inside due to its relatively bigger size. The

smaller size of the iodine allows it to move freely through the membrane while the more

complex glucose molecules cannot pass through it.

D. Osmosis

The table shows the different types of osmosis under different solvent concentrations, the

type of osmosis perceived and the net water movement in each of the conditions.

Solution Tonicity outside the Cell Cell

0.5% NaCl Isotonic

Flaccid
Distilled water Hypotonic

Turgid
10% NaCl Hypertonic

Plasmolysis
Table 2. Movement of water in the plant cells
 The Boat of Moses leaf which was penetrated by the 0.5% NaCl solution made the plant

cells become isotonic. This means that the water concentration and the solute concentration are

equal thus, the exchange of water molecules inside and outside of the cell is similar. This results

in having an unchanged shape of the plant cells. In this setup, the extracellular fluid has the same

osmolarity as the cell. The concentration inside and outside of the cell remain constant.

The Boat of Moses leaf which was penetrated by distilled water had a considerable

increase in size making it hypotonic. This means that the water concentration outside the cell is

higher compared to the water concentration inside the cell resulting into a high net movement of

water into the cell. The plant cell becomes turgid making it swell but the cell wall prevents the

cell from busting. In this setup, extracellular fluid has lower osmolarity than the fluid inside the

cell and the net movement of water will be into the cell.

The Boat of Moses leaf which was penetrated by 10% NaCl solution decreased in size

making the plant cells hypertonic. The water concentration inside the cell is higher compared to

the water concentration outside the cell having a net movement of water outside causing the cell

to shrink. In this setup, the extracellular fluid has a higher osmolarity than the cell’s cytoplasm.

Since there is a higher solute concentration outside then the water will rush out of the cell.
E. Hemolysis and crenation of the red blood cell

The table above shows the different types of osmosis under different solvent

concentrations, the type of osmosis perceived and the net water movement in each of the

conditions.

Solvents Type of Osmosis Water movement

0.5% NaCl Isotonic

Normal
Distilled water Hypotonic

Osmotic lysis
10% NaCl Hypertonic

Crenation
Table 3. Movement of water in the blood cell
 The red blood cells placed in the NaCl solution have little net osmotic movement of

water, the solution is isotonic to the RBC thus the shape and size of the cells remains the same.

The concentration of the solute inside and outside the cells is the same thus; the cells do not

shrink or swell.

The red blood cells in this setup was exposed to distilled water; it became swollen

particularly because of the higher concentration of solute found inside the cells compared to the

lower concentration outside of the cell. The water molecules penetrated the cell making the water

move inside making the RBCs become bigger in size.

The red blood cells that were placed in a 10% NaCl solution; this resulted in the RBCs

shrinking because the water moved outside of the cell because the concentration of solutes

outside the cell is higher compared to the lower concentration of the cell’s solute.

SUMMARY AND CONCLUSION

One of the most important features of the cell essential for survival is the plasma

membrane. Due to the complex structure of the cell membrane, it enables the cell to be

selectively permeable allowing various types of molecules to be transported.

The cell membrane regulates the flow of materials in and out of the cell. It possesses

certain selectivity with which it maintains a chemical balance between the cell and its

environment. There are factors that affect the permeability of the plasma membrane, such as

formalin fixation and heat. Formalin fixation decreases the permeability of the plasma membrane

by, while heat increases the movement of materials across the membrane.

Another factor that affects the movement of molecules across the membrane is the size of

the molecules. Large molecules, such as glucose and starch inside the cell are impermeable and
cannot easily pass the plasma membrane. The starch was stained by the iodine since the

amylopectin component of the starch reacts with iodine, thus the blackish-purple stain in the

collodion tube. The glucose, being a monosaccharide, does not react with the iodine having no

amylopectin chain.

The cell’s shape and size is affected by the tonicity of the solution on its environment. On

plant cell and animal cells, when the cell is exposed to a hypertonic solution, the cell shrinks and

the water inside the cell goes out. Plant and animal cells undergo plasmolysis and crenation,

respectively. On a hypotonic solution, the plant cell becomes turgid as turgid pressure increases.

However on animal cells, the animal cell inflates with the intake of water until it bursts. On an

isotonic solution, the plant cell is flaccid and the animal cell is in normal state with the equal

movement of water in maintaining the equilibrium.

The properties of the cell membrane have been crucial not only on the survival of life but

also on the practical field. Salt is added for preserving fish since the bacterial cells that causes

decomposition are killed due to the crenation of those cells. Formalin is used in the preservation

of corpses in order to prevent the water from draining out of the cells once the bodily fluids

become hypertonic upon death. The stalks of vegetables and flowers are usually immersed in

water to keep them “crunchy” since when plant cells are on a hypotonic solution, the plant cells

increases its intake of water. Through this process, plasmolysis or wilting is prevented.

The selective permeability of the plasma membrane protects the cell from its external

environment, and allows and regulates the movement of various molecules. Cellular transport is

essential in many biological processes crucial for the nutrition, reproduction, growth and

development of an organism. These processes should be further studied for more practical uses

on biotechnology.
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