The Abstract

The purpose of this lab is to determine the direction and rate of osmosis into and out of a
stimulated cells in hypotonic, hypertonic and isotonic environment. This lab will display the
factors controlling a substances direction and rate of diffusion. Lastly to show how hypertonic
solution affect the volume and integrity of plant cells.

The Objective
This lab exercise was executed from The Biology Laboratory Manual 10th Edition.
Exercise 9 Diffusion and Osmosis edited at Kean University. The purpose of this lab is to
determine the direction and rate of osmosis into and out of a stimulated cells in hypotonic,
hypertonic and isotonic environment. This lab will display the factors controlling a substances
direction and rate of diffusion. Lastly to show how hypertonic solution affect the volume and
integrity of plant cells. (Vodopich DS & Moore 2014) I predict that if one side of a membrane is
hypertonic the other side is hypotonic, osmosis, a passive transport of water molecules will travel
to the hypertonic solution. Over time the two solutions through osmosis will become isotonic.

Materials and Methods

Procedure 9.2

Our group examined one of the previously prepared agar plates under a light microscope
and noted the sizes of the halos left behind by a diffused chemical. Our group noted which
chemical had the least and most molecular mass.

Procedure 9.3
Our group obtained four pieces of strings and two pieces of water soaked dialysis tubing
each 15 cm long, and sealed one end of each tube by folding and tying it tightly. We opened the
other end of the tube by rolling in between our fingers.We filled each bag with the 10 mL of
water and three drops of phenolphthalein using a graduated cylinder and pipet and filled another
bag with 10 mL of starch suspension. For each bag, we loosely folded and secured the solution
within the bag.Our group rinsed the outside of each bag in tap water and filled a beaker with 200
mL of tap water and added 10 drops of 1 M sodium hydroxide. Finally we submerged the
dialysis bag containing phenolphthalein in the beaker and observed the results.

Procedure 9.4 & 9.5

Our group obtained eight pieces of strings and four pieces of water soaked dialysis tubing
each 15 cm long, and sealed one end of each tube by folding and tying it tightly. Then we
opened the other end of the tube by rolling in between our fingers. Then we filled each bag with
the appropriate amount of sucrose using a graduated cylinder. Afterwards we labeled each bag
accordingly by inserting a small piece of labeled paper. For each bag, we loosely folded and

secured the solution within the bag. Our group rinsed the outside of each bag in tap water, gently
blot excess liquid from the outside of the bags, and weighed each bag to the nearest 0.1g.
Our group recorded these initial weights in (table 9.1) in the first column and Placed bags B,C
and D in one largebeaker filled with 1% sucrose and placed bag A in a 250-mL beaker and filled
the beaker with 150 mL of 10% sucrose, approximately around the same time. Then we
removed the bags from the beakers at 10-min intervals for the next hour and gently blot them
dry, and weigh them to the nearest 0.1g. Afterwards we returned the bags to their respective
containers.During the 10- minute intervals the extent of water flow in each system, for each 10min interval and recorded the total weight of each bag and the content in table 9.1.

Procedure 9.7
Our group watched a video on how Procedure 9.7 was performed and observed the
results.

Procedure 9.8
We prepared a wet mount of thin layer of onion epidermis or Elodea leaf then examined the
cells. Next, we added two or more drops of 30% of NaCl to one edge of the coverslip. Our group
wicked the salt solution under the coverslip by touching a piece of absorbent paper towel to the
fluid at the opposite edge of the coverslip. We then examined the cells, the cytoplasm is no
longer pressed against the cell wall this shrinkage process is called plasmolysis.

Results
Procedure 9.2
Q2: Which would you predict would diffuse faster: a substance having a high molecular
weight or substance having a low molecular weight? Why?
The substance having a lower molecular weight would diffuse faster because smaller weighing
molecules move faster than larger/ heavier molecules.

Q3a: Considering the different molecular weight of potassium permanganate, malachite

green, and methylene blue, which should have the larger halo after the same amount of
time? Why?
Potassium permanganate should have the larger halo after time because elements with less
molecular mass diffuses faster than elements with heavier molecules and Potassium has the least
molecular mass out of these molecules.

Q3b: Do molecules stop moving when diffusion stops? Explain your answer.
No molecules are always moving as long as the temperature is above the freezing point of that
molecules.

Q4a: Describe the color changes in the two bags and their surrounding solutions.

The color change in Bag A phenolphthalein was cloudy then turned the jar and bag pink. Bag B
Sodium hydroxide initially clear was changed to a blue solution inside the bag and a red solution
outside the bag.

Q4b: For which molecules and ions does your experiment give evidence for passage
through the semipermeable membrane?
Molecules phenolphthalein, Iodine, Na(+), OH(-), all showed evidence for passing through the
semipermeable membrane. The color change within the beakers shows which molecules passed
through.

Q4c: What characteristic distinguishes those molecules and ions passing through the
membrane from those that do not pass through the membrane?
Smaller molecules pass through the membrane whereas larger molecules dont pass through the
membrane.

Q5a: Did water move across the membrane in all bags contain solutions of sugar?
Water did move across the membrane in Bag A and Bag B causing the bags to loss mass.

Q5b: In which bags did osmosis occur?

Osmosis occurred in A, C, D causing the weight of the bags to change

Q5c: A concentration gradient for water must be present in cells for osmosis to occur.
Which bag represented the steepest concentration gradient relative to its surrounding
environment?

Bag D represented the steepest concentration gradient. There was 20% sucrose in dialysis.

Q5d: The steepest gradient should result in the highest rate of diffusion. Examine the data
in Table 9.1 for Change in Weight during the 10 -20 minute interval. Did the greatest
change in weight occur in cells with the steepest concentration gradients? Why or Why
not?
Our data did not support the greatest change in weight occur in cells with the steepest
concentration. Our results showed only one gram was increased from the bags highest in weight.
The best weight occurs in cells that are similar in concentration to the environment

Procedure 9.8
Q9a. Why did the plant cells plasmolyze when immersed in a hypertonic solution?
The cells lost water due to osmosis, the water wants to travel to the place with the highest
solute and this was outside of the cell.

Q9b. What can you conclude about the permeability of the cell membrane (i.e., the membrane
surrounding the cytoplasm) and vacuolar membrane (the membrane surrounding the vacuole) to
water?

The cell membrane was permeable to H2O, the central vacuole shrunk.

Procedure 9.4: Table 9.1

Inside of Bag-vs.
solution in beaker

0 minutes 10minutes
Total
Total
Weight
Weight

20
minutes
Total
Weight

30
minutes
Total
Weight

40
minutes
Total
Weight

11g

11g

11g

11g

11g

10g

9g

9g

8g

8g

11g

12g

12g

12g

12g

11g

12g

12g

12g

12g

Hypotonic

B
Isotonic

C
Hypertonic

D
Hypertonic

Changes in Weight of Dialysis Bags Used as Cellular Models

11
10

0 minutes

Discussion

12
11

12
11

12
11

12
11

20 minutes

30 minutes

40 minutes

10 minutes

Hypotonic A

Isotonic B

Hypertonic C

Hypertonic D

The data I obtained refuted my initial hypothesis. For Procedure 9.8 Bag A is supposed to
decrease by Law of osmosis. Bag A possibly could have weighed the same through each
recording from a rounding error of the scale. Bag A could have been tied to tightly therefore no
allowing enough solution to permeate through. Bag B is supposed to maintain the same weight
since it is isotonic to its environment; Because Bag B was submerged in a beaker with other
dialysis bags that are not isotonic instead hypertonic, the environment changed its molecules
concentration thus leaving Bag B more hypotonic then its environment. By Law of Diffusion we
observed Bag B losing mass while Bag C and D were becoming more isotonic to its
environment. ( Reece, J. B., Urry, L. A., Cain, M. L., Wasserman, S. A., Minorsky, P. V., Jackson, R.,
& Campbell, N. A.)

Conclusion
The purpose of this lab was to determine the direction and rate of the cells
across a concentration gradient. Bag A shrank as we predicted it would. This
happened because the 10% sucrose solution in the beaker contained more
solutes than the 1% sucrose solution inside the bag. The weight of the bag
remained the same with each 10 minutes interval. Bag B decreased after the
first 10 minutes interval and remained the same. The initial weight of the
bag was 10grams after the first 10 minutes it weighed 9grams the bag
showed no change in mass for the remainder of the time. Bags C and D had
the same results in terms of swelling. These two bags would show an
increase in mass because of sucrose solution was greater inside the bag at
10% than outside the bag at 1%. Our hypothesis stated if one side of a

membrane is hypertonic, the other is hypotonic, than through osmosis, a

passive transport of water will travel to the hypertonic solution. Our data and
hypothesis did support our conclusion.

Citations

1. Reece, Jane B., Lisa A. Urry, Michael L. Cain, Steven Alexander Wasserman, Peter V.
Minorsky, Rob Jackson, and Neil A. Campbell. "Chapter 7 Membrane Structure and
Function. Chapter 3 Water and Life." Campbell Biology.
2. Bio 1300 Gen BIO Lab Exercise Diffusion & Osmosis Vodopich DS & Moore R
(2014) The Biology Laboratory Manual 10th Edition. Exercise 9 Diffusion and Osmosis
3. Hemolysis video. (n.d.). Retrieved from https://www.youtube.com/watch?
v=65J6JN6j2fA