Exercise 3

MEMBRANE TRANSPORT

A cell continuously exchanges substances with its surrounding environment to maintain homeostasis. Such
process is regulated by the plasma membrane, a semi-permeable structure that allows the entry and exit of
some materials into and out of the cell more easily than the others.

Membrane transport can be categorized as either passive or active based on the use of energy. Passive
transport works without expending energy. It is powered by concentration gradient, that is, molecules move
from an area of high concentration to that of lower concentration. Some forms of this transport include simple
diffusion and osmosis. Diffusion is simply the random movement of molecules down their concentration
gradient. Osmosis, however, specifically describes the diffusion of water molecules thru a selectively permeable
membrane.

On the contrary, active transport makes use of energy and moves against the concentration gradient. Known
forms of this process include protein pumps, endocytosis and exocytosis.

OBJECTIVES
At the end of the exercise, the students are able to:
1. identify some factors that would affect the diffusion of substances; and
2. state the effects of tonicity in an osmotic process.

MATERIALS
Potassium Permanganate (KMnO4) Graduated Cylinder Medicine Dropper
Methyl Orange Test Tube Yarn
Malachite Green Beaker Paper Towel
Methylene Blue Petri Plate Sharp Razor Blade
5% Sodium Chloride (NaCl) Stirring Rod Bixa orellana (Atsuete)
5% Sucrose Solution Test Tube Rack Tradescantia spathacea (Bangka-bangkaan)
25% Sucrose Solution Ruler Allium cepa (Onion Bulb)
Agar or Gelatin Cover Slip Sugar Beet
Vegetable Oil Microscope Slide
Longganisa Skin Microscope

PROCEDURES
A. Diffusion of Selected Plant Pigments
1. Label 4 test tubes as #1, #2, #3 and #4 respectively.
2. Put 1 gram of atsuete seeds into the test tubes and add the following as instructed:
a) test tube 1: 10mL distilled water
b) test tube 2: 10mL distilled water and place in boiling water bath
c) test tube 3: 10mL vegetable oil
d) test tube 4: 10mL vegetable oil and place in boiling water bath
3. After 30 minutes, shake the tubes and compare the relative color intensities using the symbols +, ++, +
++, ++++ (referring to an increasing color intensity).
4. Repeat the procedure using 0.5 grams of sugar beets.
5. Record your observations.
B. Diffusion in Solids
 1. Label your agar or gelatin plates as follows:
a. Plate 1: Potassium Permanganate
b. Plate 2: Methyl Orange
c. Plate 3: Malachite Green
d. Plate 4: Methylene Blue
2. Place grains (or three drops) of the abovementioned substances on their respective plates at the same
time and allow them to diffuse.
3. At 15-minute intervals, measure (in centimeters) the radius of the halos of color with a ruler.
4. Record your observations.

C. Osmosis in Artificial Cells

1. Label three 250mL beakers as #1, #2 and #3 and add the following as instructed:
a) beaker 1: 150mL distilled water
b) beaker 2: 150mL 5% sucrose solution
c) beaker 3: 150mL 25% sucrose solution
2. Fold and tie off one end of the longganisa skin. Open its other end by rolling it between your fingers.
3. Fill each of the longganisa skin with 10mL 5% sucrose solution.
4. Fold and tie off the open end. Blot off any excess solution and weigh each bag. Record the initial
weights.
5. Place the longganisa skin to their respective beakers at the same time. Take note of the time when you
placed the bags in the beakers.
6. After one hour, remove the bags, blot them dry and weigh again. Record their final weight.
7. Compute for the percent change in weight (%ΔW) per set-up as follows:

final weight-initial weight

%ΔW= *100
initial weight

D. Osmosis in Plant Cells

1. Strip off thin sections of the lower epidermis of bangka-bangkaan (the side with the violet color).
2. Place a section in a microscope slide and add a drop of distilled water.
3. Cover the wet mount and examine the cells under LPO.
4. Sketch a cell in your worksheet. Label the visible parts.
5. Without moving the slide, draw off the water with a piece of paper towel and add a drop of 5% sucrose
solution. Examine again the cells under LPO and note the change in any one cell.
6. Make a sketch of the cell. Label the visible parts.
7. Repeat the process using onion bulb.

REFERENCES
1. Ragragio, E. M. and K. V. Evangelista. 2004. Laboratory Manual in Plant Physiology. University of the
Philippines-Manila.
2. http://www.saddleback.edu/faculty/steh/bio3afolder/bio3atransportlab.pdf
3. www.usi.edu/.../Lab%20Ex%20Membranes%20and%20Cell%20Transport%20Use.ppt
4. http://www.zoology.ubc.ca/~harris/files/permL.doc

Name:_____________________________________________________ Score:________________________
Course & Year:______________________________________________ Date Performed:________________

Exercise 3
MEMBRANE TRANSPORT

A. Diffusion of Selected Plant Pigments

Set-up Color Intensities (+, ++, +++, ++++)
Atsuete Sugar Beet
Test Tube 1: 10mL distilled water
Test Tube 2: 10mL heated distilled water
Test Tube 3: 10mL vegetable oil
Test Tube 4: 10mL heated vegetable oil

1. Which set-up had the most intense color for atsuete? for sugar beet? Explain why.
__________________________________________________________________________________
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2. What is the role of heat in the diffusion process?
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B. Diffusion in Solids
Time Radius of Diffusion Halo (cm)
(min) Potassium Permanganate Methyl Orange Malachite Green Methylene Blue
15
30
45
60
75
90

1. Which substance reached the largest radius after 90 minutes?

__________________________________________________________________________________
2. What factor could possibly allow your answer in #1 to reach such radius? Explain why.
__________________________________________________________________________________
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3. In general terms, describe the relationship of your answer in #2 and the radius of diffusion halo of each
substance you used in this specific experiment.
__________________________________________________________________________________
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C. Osmosis in Artificial Cells

 Set-up Initial Weight Final Weight % Change in Weight Tonicity
(go) (g) (g) (solution to bag)
Bag in Distilled Water
Bag in 5% Sucrose
Bag in 25% Sucrose
Note: bag contains 5% sucrose solution

1. What is the direction of movement of water in the following set-ups:

a) Beaker 1: _________________________________
b) Beaker 2: _________________________________
c) Beaker 3: _________________________________
2. How would raising the concentration of sucrose in the bag to 10% affect the movement of water
molecules in beakers 2 and 3? Explain why.
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
__________________________________________________________________________________
3. Based on your observations, is the longganisa skin permeable to sucrose? Why?
__________________________________________________________________________________
__________________________________________________________________________________
4. What is dialysis? How is it different from osmosis?
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D. Osmosis in Plant Cells

Magnification: __________ Magnification: __________

Figure 1. Cell of bangka-bangkaan before (left) and after (right) exposure to 5% sucrose solution.
Magnification: __________ Magnification: __________

Figure 2. Cell of onion bulb before (left) and after (right) exposure to 5% sucrose solution.

1. Based on your observations, differentiate turgidity and plasmolysis in plant cells.

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2. Which part of the plant cell collapsed after exposure to 5% solution? What is the function of this
structure?
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3. What is effective osmotic pressure? How does it relate to cellular osmosis?
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