Diffusion Through the Cell Membrane and Osmosis 1

Diffusion Through the Cell Membrane and Osmosis

Kailey McSteen
Honors Biology Period 5
Cardinal Wuerl North Catholic High School
April 30, 2018
 Diffusion Through the Cell Membrane and Osmosis 2

Introduction

Cell transport is a topic that is often misunderstood by the general public. There are two

main types of cell transport, and these include active transport and passive transport. The focus

of the lab is passive transport. Passive transport is when molecules move from a high area of

concentration to a low area of concentration across a cell membrane. Passive transport happens

naturally, and it does not require the cell to exert any energy or ATP in order for the movement

to occur (OpenStax College). The cell membrane does not let every substance pass through it,

and this means that it is selectively permeable. Specifically, selectively permeable means that the

cell membrane allows some substances to pass through it, while other substances can not pass

through. If the cell membrane were not selectively permeable, then the cell would experience

problems. The cell would most likely be destroyed because it would not be able to sustain itself

any longer (OpenStax College).

Osmosis is relevant to this lab because it is the movement of water from a high

concentration to a low concentration across a cell membrane. Osmosis is a type of passive

transport that utilizes channel proteins called aquaporins in order for the process to occur.

Aquaporins are always open, so this means that there is always a net movement of water across

the cell membrane. Osmosis uses aquaporins to assist in the process because water is polar, and

water would not be able to pass through the non-polar fatty acid tails of the cell membrane

without the help of a channel proteins. Osmosis is specific to water (Biggs, et al., 2012). There

are three different types of osmotic environments that the cell can be placed into. The first type

of osmotic environment is a hypotonic environment. In a hypotonic environment, there is a

higher concentration of pure water outside the cell, and there is a lower concentration of pure

water inside the cell. This is a type of passive transport, so the water will move inside the cell
 Diffusion Through the Cell Membrane and Osmosis 3

because it is going from a high concentration to a low concentration. The cell could burst

because of all the water that is rushing in. The water is moving with the concentration gradient

(Biggs, et al., 2012). The second type of osmotic environment is an isotonic environment. In an

isotonic environment, there is same concentration of pure water inside the cell as there is outside

of the cell. The movement of water outside the cell is balanced with the movement of water

inside the cell. In an isotonic environment, the concentration of pure water inside the cell and

outside the cell would be very close to being the exact same amount, but there might be slightly

more or less inside or outside the cell because of the net movement of water. The size of the cell

will remain constant (OpenStax College). The third type of osmotic environment is a hypertonic

environment. In a hypertonic environment, there is a higher concentration of pure water inside

the cell than there is outside the cell, so the water would be moving outside of the cell. This

could cause the cell to shrivel. In a hypertonic environment, the water is moving with the

concentration gradient (Biggs, et al., 2012).

It is important to understand the process of osmosis because it can be applied to real life

situations. For example, in the grocery store, the vegetables are usually sprayed with water. The

vegetables are sprayed with water because this places them into a hypotonic environment. This

keeps the vegetables fresh, and it prevents them from shriveling up. Another example of this

when meat is salted in order for it to be preserved. This puts the meat into a hypertonic

environment. The water would move outside, which would help to preserve the meat.

Understanding osmosis would be helpful because knowing when the cell swells and shrinks

could be useful in multiple situations (Diffusion Through Cell Membrane Lab Guide, 2018).

Dialysis tubing is used to simulate a cell membrane in the lab. Dialysis tubing is made of

selectively permeable cellulose, and the tubing also has microscopic pores. This means that
 Diffusion Through the Cell Membrane and Osmosis 4

dialysis tubing is accurate in helping to represent the movement of substances across the cell

membrane (Hanley, Herting, & Smith, 2013). There are various purposes for completing this lab.

The first purpose of the lab is to see the different effects of osmosis in the different osmotic

environments. The second purpose is to create a model of what an actual cell membrane would

be like. The third purpose is to see how the rate of osmosis differs with different concentration

gradients. The fourth purpose is to see how the rate of osmosis changes when you get close to

equilibrium.

For the setup of the lab, there are 4 beakers that contain 200 mL of pure water, and there is

one beaker that contains 200 mL of an 60% glucose solution. Setup 1 represents an isotonic

environment because the dialysis tubing if filled with pure water, and it is placed in a beaker of

pure water. Setup 2 represents a hypotonic environment because the dialysis tubing is filled with

a 20% glucose solution, and it is placed in a beaker of pure water. Setup 3 represents a hypotonic

environment because the dialysis tubing is filled with a 40% glucose solution, and it is placed in

a beaker of pure water. Setup 4 represents a hypotonic environment because the dialysis tubing is

filled with a 60% glucose solution, and it is placed in a beaker of pure water. Setup 5 and 6

represents both a hypertonic environment and a hypotonic environment. This is the case because

the beaker has two different dialysis tubes placed in a 60% glucose solution. The first dialysis

tube is filled with pure water, so this is the hypertonic environment. The second dialysis tube is

filled with an 80% glucose solution, so this is the hypotonic environment (Diffusion Through

Cell Membrane Lab Guide, 2018).

The dependent variable for part I is the mass change of the dialysis tubing. The

independent variable for part I is the osmotic solution, and this is what you change from group to

group in order to see the mass change. The dependent variable for part II is the color change. The
 Diffusion Through the Cell Membrane and Osmosis 5

independent variable for part II is the location of the starch. The constants for part I are the

temperature of the solution, the starting amount of liquid in each dialysis tube, the starting

amount of liquid in each beaker, the time each dialysis tube was left in the liquid, how the

dialysis tube was folded and tied, and how the tubes were dried before they were weighed. The

control group for part I is setup 1, water in water, and the experimental groups for part I are the

other 5 setups. The constants for part II are the amount of iodine (20 drops), half a spoonful of

starch, washing the simulated cell thoroughly before putting it in the solution, and the amount of

water in the beaker. The control group for part II is the original setup with yellow water and

white starch in a simulated cell. The experimental group for part II is the after with clear water

and dark purple/ black color inside the simulated cell. There are 6 hypotheses for part I: If a

dialysis tube filled with 5 mL of water is placed in a beaker of 200 mL of water, then the dialysis

tube will maintain the same mass due to its isotonic environment. If a dialysis tube filled with 5

mL of a 20% glucose solution is placed in a beaker if 200 mL of water, then the dialysis tube

will gain mass due to its hypotonic environment. If a dialysis tube filled with 5 mL of a 40%

glucose solution is placed in a beaker of 200 mL of water, then the dialysis tube will gain mass

due to its hypotonic environment. If a dialysis tube filled with 5 mL of a 60% glucose solution is

placed in a beaker if 200 mL of water, then the dialysis tube will gain mass due to its hypotonic

environment. If a dialysis tube filled with 5 mL of water is placed in a beaker of 200 mL of a

60% glucose solution, then the dialysis tube will lose mass due to its hypertonic environment. If

a dialysis tube filled with 5 mL of a 80% glucose solution is placed in a beaker of 200 mL of a

60% glucose solution, then the dialysis tube will gain mass due to its hypotonic environment.

The hypothesis for part II is: If a dialysis tube is filled half way with starch and it placed in a

solution with iodine, then the color of the dialysis tubing will be blue/black after 15 minutes.
 Diffusion Through the Cell Membrane and Osmosis 6

Materials

 5 beakers for part I (200 mL)

 1 beaker for part II (200 mL)

 20% glucose solution

 40% glucose solution

 60% glucose solution

 80% glucose solution

 Pure water solution

 Dialysis tubing

 Scale (mg)

 Strings

 Paper towels

 Timer

 5 Pipets

 Iodine

 Potato Starch

 5 Graduated cylinders (5 mL)

 Scissors

 Plastic spoons

Procedures

For part I of the lab, the goal is to see the effect of concentration on the rate of diffusion.

First, take 6 pieces of dialysis tubing that have been soaking in water. Take the dialysis tubing
 Diffusion Through the Cell Membrane and Osmosis 7

out of the water, and fold about 1 inch from the bottom of the dialysis tubing in two ways. The

first fold is a hamburger style fold, and the second fold is a hot dog style fold. After the dialysis

tubing is folded, take the string and tie a very tight knot around the folded part of the dialysis

tubing. Then, cut the excess string. Repeat this process with all 6 dialysis tubes (Diffusion

Through Cell Membrane Lab Guide, 2018). The first dialysis tube is filled with 5 mL of pure

water. The second dialysis tube is filled with 5mL of a 20% glucose solution. The third dialysis

tube is filled with 5mL of a 40% glucose solution. The fourth dialysis tube is filled with 5mL of

a 60% glucose solution. The fifth dialysis tube is filled with 5 mL of pure water. The sixth

dialysis tube is filled with 5mL of a 80% glucose solution. After each dialysis tube is filled,

repeat the same folding and tying process to the other end of each dialysis tube until all 6 dialysis

tubes are tied on both ends (Diffusion Through Cell Membrane Lab Guide, 2018). Get 6 paper

towels and label the paper towels according to the type of solution that is in each dialysis tubing.

Place each dialysis tube on the paper towel that is labeled for it. Then, get 4 beakers and fill them

up with 200 mL of water. Fill one beaker up with 200 mL of a 60% glucose solution (Diffusion

Through Cell Membrane Lab Guide, 2018). Get the mass of each dialysis tube using a scale.

Record the starting mass of each dialysis tube in a data table. Next, place each dialysis tube in a

separate beaker with pure water. The one dialysis tube filled with pure water, and the other

dialysis tube filled with a 80% glucose solution need to be placed in the beaker with a 60%

glucose solution. These two dialysis tubes will be placed in the same beaker. All of the dialysis

tubes need to be put in the beakers at the same time. When the dialysis tubes are placed inside

the beakers, start a timer. At the end of each 3, 6, and 9 minute period, remove the dialysis tubes

from the beakers and place them on the labeled paper towels. Dry each dialysis tube by rolling it

on the paper towel. Then, weigh each dialysis tube on the scale in either grams or milligrams.
 Diffusion Through the Cell Membrane and Osmosis 8

Record the masses in a data table. When finished weighing the dialysis tubes, return the bags to

the correct beakers at the same time (Diffusion Through Cell Membrane Lab Guide, 2018).

For part II of the lab, the goal is to see if certain substances can pass in or out of a cell

model. First, take one dialysis tube and use the folding and tying method from part I of the lab to

tie one end of the dialysis tube. Once the one end of the dialysis tube is completely tied, add

roughly one teaspoon of potato starch to the dialysis tube. Then, add roughly 5 mL of pure water

to the dialysis tube that already has the potato starch in it. Use the folding and tying method to tie

the other end of the dialysis tube (Diffusion Through Cell Membrane Lab Guide, 2018). Once

the dialysis tube is completely tied, shake the dialysis tube to make sure the water and the potato

starch are mixed up. Next, rinse the dialysis tube off with water to clean off any spills. Pat the

dialysis tube with a paper towel to make sure that it is dry. Set the dialysis tube to the side for a

short period of time. Then, fill a beaker roughly half full of water, and add around 10 drops of

iodine to the beaker. Place the dialysis tube in the iodine water. Record the initial color of the

dialysis tube, which is the model cell, in a data table, and also record the initial color of the

beaker. Take the dialysis tube out of the beaker after 15 minutes. Note if there is any change in

color in the beaker or the dialysis tube. Record any changes in color in the data table that was

previously made (Diffusion Through Cell Membrane Lab Guide, 2018).

Results

For part I of the lab, there are various results. A class average was taken for the change in

mass of the dialysis tubes for 0-3 minutes, 3-6 minutes, and 6-9 minutes. Setup 1 was the dialysis

tube filled with 5 mL of water that was placed in a beaker of 200 mL of water. The average

change in mass of the dialysis tube filled with water placed in the beaker of water for 0-3

minutes was 208 mg. The average change in mass for the dialysis tube filled with water placed in
 Diffusion Through the Cell Membrane and Osmosis 9

water for 3-6 minutes was 83 mg. The average change in mass for the dialysis tube filled with

water placed in water for 6-9 minutes was -41 grams. For this setup, there was increase in mass

at 3,6, and 9 minutes, but as time went on, the increase in mass was not as high. Refer to figure

1. Setup 2 was the dialysis tube filled with a 20% glucose solution placed in a beaker of 200 mL

of water. The average change in mass of the dialysis tube filled with a 20% glucose solution

placed in a beaker of water for 0-3 minutes was 317 mg. The average change in mass of the

dialysis tube filled with a 20% glucose solution placed in a beaker of water for 3-6 minutes was

217 mg. The average change in mass of the dialysis tube filled with a 20% glucose solution

placed in a beaker of water for 6-9 minutes was 167 mg. For this setup, there was increase in

mass at 3,6, and 9 minutes, but as time went on, the increase in mass was not as high. Refer to

figure 1. Setup 3 was the dialysis tube filled with a 40% glucose solution placed in a beaker of

200 mL of water. The average change in mass of the dialysis tube filled with a 40% glucose

solution placed in a beaker of water for 0-3 minutes was 408 mg. The average change in mass of

the dialysis tube filled with a 40% glucose solution placed in a beaker of water for 3-6 minutes

was 392 mg. The average change in mass of the dialysis tube filled with a 40% glucose solution

placed in a beaker of water for 6-9 minutes was 308 mg. For this setup, there was increase in

mass at 3,6, and 9 minutes, but as time went on, the increase in mass was not as high. Refer to

figure 1. Setup 4 was the dialysis tube filled with a 60% glucose solution placed in a beaker of

200 mL of water. The average change in mass of the dialysis tube filled with a 60% glucose

solution placed in a beaker of water for 0-3 minutes was 567 mg. The average change in mass of

the dialysis tube filled with a 60% glucose solution placed in a beaker of water for 3-6 minutes

was 442 mg. The average change in mass of the dialysis tube filled with a 60% glucose solution

placed in a beaker of water for 6-9 minutes was 400 mg. For this setup, there was increase in
 Diffusion Through the Cell Membrane and Osmosis 10

mass at 3,6, and 9 minutes, but as time went on, the increase in mass was not as high. Refer to

figure 1. Setup 5 was the dialysis tube filled with pure water placed in a beaker with 200 mL of a

60% glucose solution. The average change in mass of the dialysis tube filled with pure water

placed in a beaker of a 60% glucose solution for 0-3 minutes was -150 mg. The average change

in mass of the dialysis tube filled with pure water placed in a beaker of a 60% glucose solution

for 3-6 minutes was -383 mg. The average change in mass of the dialysis tube filled with pure

water placed in a beaker of a 60% glucose solution for 6-9 minutes was -250 mg. For this setup,

there was a loss of mass at 3,6, and 9 minutes, but the most loss of mass was from 3-6 minutes.

Refer to figure 1. Setup 6 was the dialysis tube filled with a 80% glucose solution placed in a

beaker with 200 mL of a 60% glucose solution. The average change in mass of the dialysis tube

filled with a 80% glucose solution placed in a beaker of a 60% glucose solution for 0-3 minutes

was 241 mg. The average change in mass of the dialysis tube filled with a 80% glucose solution

placed in a beaker of a 60% glucose solution for 3-6 minutes was 75 mg. The average change in

mass of the dialysis tube filled with a 80% glucose solution placed in a beaker of a 60% glucose

solution for 6-9 minutes was 83 mg. For this setup, there was an increase in mass at 3,6, and 9

minutes, but the highest increase in mass was from 0-3 minutes. Refer to figure 1. In order to

graph this data to show mass changing over time, the masses at 0,3, 6, and 9 minutes were

needed. All starting masses were placed at 0 milligrams. In order to calculate the mass at 3,6, and

9 minutes the changes in mass were needed. Refer to table 1.

The table below shows the change in mass of the dialysis tubes in the different osmotic

solutions. The measurements of the table are all in milligrams. The measurements in the table are

class averages taken from the results of each individual group in the different class periods. The
 Diffusion Through the Cell Membrane and Osmosis 11

graph below is representing the data in the table in a visual form. There are six series on the

graph that represent all of the dialysis tubes in the various osmotic environments.

Table 1: Changes in Mass of the Dialysis Tubes in Milligrams in Different Solutions

Time Water in 20 % in 40% in 60% in Water in 80% in
Water Water Water Water 60% 60%
0 0 0 0 0 0 0
3 208 317 408 567 -150 241
6 291 534 800 1009 -533 316
9 249 701 1108 1409 -783 399
Description: This data was collected by multiple groups in many classes. Since each group
started with a different initial mass of their dialysis tubes, mass changes from 0-3 min, 3-6 min,
and 6-9 min were recorded and averaged between the groups. In order to represent mass change
on a graph, masses are needed for 0, 3, 6, and 9 min. To have consistent data, the starting masses
at 0 minutes of the dialysis tubes were 0 mg. The masses at 3 minutes are the changes in mass
from 0-3 minutes. The mass at 6 minutes was calculated by adding the change in mass from 3-6
minutes to the previous mass. The mass at 9 minutes was calculated by adding the change in
mass from 6-9 minutes to the previous mass.

Mass in Miligrams vs. Time in Minutes

2000

1500
Water in water
1000 20% in water
Mass (mg)

500 40% in water

60% in water
0
0 3 6 9 Water in 60%
-500
80% in 60%
-1000
Time (minutes)

Figure 1: Changes in Mass of the Dialysis Tubes Over Time

Description: In the graph above, there are six series representing the different dialysis tubes.
There is the dialysis tube filled with water in water, the dialysis tube filled with a 20% glucose
solution in water, the dialysis tube filled with a 40% glucose solution in water, the dialysis tube
filled with a 60% glucose solution in water, the dialysis tube filled with water in a 60% glucose
solution, and the dialysis tube filled with a 80% glucose solution in a 60% glucose solution. The
Y axis is showing the mass of the dialysis tubes in milligrams. The X axis is showing the time in
minutes that the dialysis tubes spent in the different solutions. The series are each represented by
a different color, and the key shows which color corresponds to which series. This graph shows a
 Diffusion Through the Cell Membrane and Osmosis 12

visual representation of the change in mass for each dialysis tube, and this allows comparisons to
be made.

For part II of the lab, the starting color of the solution in the dialysis tube was white, and the

starting color for the solution in the beaker was yellow. The color of the dialysis tube after it was

submerged in the iodine solution for 15 minutes was blue/black. The color of beaker was a very

pale yellow or almost clear.

Discussion

For setup 1, the water in water, the dialysis tube initially gained weight, but the weight stayed

relatively around the same during the entire 9 minutes. The reason that this dialysis tube

maintained around the same weight was because it was placed an isotonic environment. In an

isotonic environment, there is the same concentration of pure water both inside and outside of the

cell. The reason that the weight of the cell would fluctuate a little bit is due to aquaporins, which

are always open. This causes a net movement of water, and this concept was also applied to the

dialysis tube in the lab. For setup 2, the 20% glucose solution in water, the dialysis tube gained

weight because it was placed in a hypotonic environment. There was a higher concentration of

pure water outside the cell, so water was rushing in. For setup 3, the 40% glucose solution in

water, the dialysis tube gained weight because it was placed in a hypotonic environment. For

setup 4, the 60% glucose solution in water, the dialysis tube gained weight because it was placed

in a hypotonic environment. For setup 5, the water in a 60% glucose solution, the dialysis tube

lost weight because it was placed in a hypertonic environment. In a hypertonic environment,

there is a higher concentration of pure water inside the cell, so water is rushing out. This concept

was applied to the dialysis tube in the lab. For setup 6, the 80% glucose solution in the 60%

glucose solution, the dialysis tube gained weight because it was placed in a hypotonic
 Diffusion Through the Cell Membrane and Osmosis 13

environment. The rate of osmosis decreases as the dialysis tubes get closer to equilibrium. This

concept can be applied to all of the setups in the lab. When there is a higher concentration

gradient, that means there is very big difference in the concentration of pure water. This means

that the rate of osmosis would increase because the cell would be farther away from reaching

equilibrium. When there is a low concentration gradient, that means there is small difference in

the concentration of pure water. This means that the rate of osmosis would decrease because the

cell would be closer to reaching equilibrium. The dialysis tube filled with an 80% glucose

solution placed in a 60% glucose solution did not gain as much weight from 0-3 minutes as the

dialysis tube filled with a 20% glucose solution placed in pure water partly due to human error.

The data that was used in the lab was averages taken from multiple different class periods. This

meant that outliers could have affected the average taken for the dialysis tube filled with an 80%

glucose solution placed in a 60% glucose solution. Even though the 80/60 dialysis tube was

placed in a beaker with a dialysis tube filled with pure water, this did not affect the change in

mass because one dialysis tube was in a hypertonic environment and the other dialysis tube was

in a hypotonic environment. Water was rushing out of the dialysis tube filled with pure water

placed in the 60% glucose solution, and this would increase the concentration gradient for the

other dialysis tube inside the beaker. The rate of osmosis should have increased due to the

increase in the concentration gradient, but this was not the case for the 80/60 dialysis tube. The

same change in mass was expected for the 80/60 dialysis tube and the 20/0 dialysis tube because

both of those have a 20% concentration gradient, but that did not happen.

For part II of the lab, the inside of the dialysis tube turned blue because the dialysis tube was

permeable to the iodine solution. The dialysis tubing was not permeable to starch. If the dialysis

tube was permeable to starch, then the color inside the beaker would have been blue. If the
 Diffusion Through the Cell Membrane and Osmosis 14

dialysis tube was not permeable to anything, then there would have been no color change at all.

The starch stayed inside the dialysis tube, and it did not pass through the dialysis tube (simulated

cell) into the beaker. One source of error was that the temperature of the solution in the beaker

was different for reach setup. Another source of error was that the dialysis tubes were not

thoroughly dried before they were placed back into the beakers. The third source of error is that

some of the dialysis tubes were not tied as tightly as they could have been tied. The final source

of error is that the scale was not dried off completely before weighing all of the dialysis tubes.

One change to make the lab better is letting the dialysis tubes sit in the different solutions for

longer periods of time than 3, 6, and 9 minutes. Letting them sit longer in the solutions would

provide more accurate results. In conclusion, all 6 hypotheses for part I were supported, and the

hypothesis for part II was also supported.

 Diffusion Through the Cell Membrane and Osmosis 15

References

Biggs, A., Hagins, W., Holliday, W., Kapicka, C., Lundgren, L., MacKenzie, A., . . . Zike, D.

(2012). Biology. Columbus: McGraw-Hill.

Diffusion Through Cell Membrane Lab Guide. (2018, April). Diffusion Through Cell

Membranes.

Hanley, P., Herting, K., & Smith, S. (2013, April 18). Separation Characteristics of Dialysis

Membranes. Retrieved from Thermofisher:

https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-biology-

learning-center/protein-biology-resource-library/protein-biology-application-

notes/separation-characteristics-dialysis-membranes.html

OpenStax College. (2014, March 28). Passive Transport. Retrieved from the OpenStax-CNX

Web site: https://legacy.cnx.org/content/m44417/1.12/