PLASMA MEMBRANCES & DIFFUSION

PHED 2516 – LAB 2

DATA COLLECTION WORKSHEET
Open the PhysioEx 10.0 software package using the link and login below. Download Adobe Flash Player
if necessary (a link will be on the page that opens).
https://media.pearsoncmg.com/bc/bc_0media_ap/physioex/10/
Login = phed_lab
Password = PhedLab1

Experiment #1 – Simple Diffusion

1) In the PhysioEx 10.0 main menu, click Exercise 1: Cell Transport Mechanisms and Permeability,
then click Activity 1: Simulating Dialysis (Simple Diffusion). Once the activity opens click on the
Experiment tab (in the dropdown menu of the 3 bar icon in the top left corner of the webpage).
Ignore the instructions at the top. We are just using the software to simulate our own experiment.
2) On the left side of the simulation, click the + sign beside each of the Na+/Cl-, Urea, Albumin and
Glucose concentration indicators until they all read 5mM. Then click the Dispense icon. Note: mM
is the abbreviation for millimoles.
3) On the right side of the simulation, click the Deionised Water icon and then the Dispense icon.
4) Click and hold on the 20 MWCO membrane tube to the far right. Drag the tube to the middle of
the beakers at the top of the simulation and release it. Note: MWCO stands for Molecular weight
cut-off and can be thought of in terms of pore size where a larger MWCO number indicates larger
pore sizes in the membrane.
5) Click the Start icon and observe the concentration levels displayed beside each beaker.
6) Click Record Data.
7) Record the final concentrations of Na +/Cl-, Urea, Albumin and Glucose in the left and right beakers
(using the numbers beside the beakers) along with their diffusion rates (from the recorded data)
into Table 1 below.
8) Drag the 20 MWCO membrane back to its original location and click both Flush icons.
9) Repeat for the remaining 3 membranes.
10) Answer the following questions as you work through the experiment.
 Which solutes where able to diffuse from the left to the right beaker?
 Did they diffuse at the same rate? Why?
 Why where some solutes unable to move from the left beaker to the right? How could you
move those solutes across the membrane?
11) Return to the PhysioEx 10.0 main menu.

PHED 2516 – Lab 2: Experiments Worksheet Page 1 of 10

Table 1: Simple diffusion results.
Initial Initial Final Final
MWCO Diffusion
Concentration Concentration Concentration Concentration
Trial Solutes Rate
(g/mol) Left Beaker Right Beaker Left Beaker Right Beaker
(mM/sec)
(mM) (mM) (mM) (mM)

Na+ / Cl- 20 5 0 5 0 0
Urea 20 5 0 5 0 0
1
Albumin 20 5 0 5 0 0
Glucose 20 5 0 5 0 0
+ -
Na / Cl 50 5 0 2.5 2.5 0.0093
Urea 50 5 0 0 0
2 0
Albumin 50 5 0 5 0 0
Glucose 50 5 0 5 0 0
+ -
Na / Cl 100 5 0 2.5 0.0093
2.5
Urea 100 5 0 2.5 2.5 0.0056
3
Albumin 100 5 0 0 0
0
Glucose 100 5 0 0 0 0
Na+ / Cl- 200 5 0 2.5 2.5 0.0093
Urea 200 5 0 2.5 2.5 0.0056
4
Albumin 200 5 0 0 0
0
Glucose 200 5 0 2.5 0.0024
2.5

PHED 2516 – Lab 2: Experiments Worksheet Page 2 of 10

Experiment #2 – Facilitated Diffusion
1) In the PhysioEx 10.0 main menu, click Exercise 1: Cell Transport Mechanisms and Permeability,
then click Activity 2: Simulating Facilitated Diffusion. Once the activity opens click on the
Experiment tab. Again, ignore the instructions at the top.
2) On the left side of the simulation, click the + sign beside the Glucose concentration indicator until
it reads 2mM. Then click the Dispense icon.
3) On the right side of the simulation, click the Deionised Water icon and then the Dispense icon.
4) On the far right side of the simulation, click the + sign below the Glucose Carriers indicator until
500 glucose carriers are selected. Then click Build Membrane.
5) Click and hold on the glucose carrier membrane tube. Drag the tube to the middle of the beakers
at the top of the display and release it.
6) Click the Start icon and observe the concentration levels displayed beside each beaker.
7) Click Record Data.
8) Record the final concentrations of Glucose in the left and right beakers (using the numbers beside
the beakers) along with its diffusion rate (from the recorded data) into Table 2 below.
9) Drag the glucose carrier membrane back to its original location and click both Flush icons.
10) Repeat using a membrane with 900 glucose carriers.
11) Repeat the 500 and 900 glucose membrane trials with 8mM of Glucose in the left beaker.
12) Repeat the last trial 900 glucose membrane trial with the same 8mM of Glucose of glucose in both
beakers.
13) Answer the following questions as you work through the experiment.
 Was the rate of diffusion increased or decreased when you increased the number of glucose
carriers? Why?
 What happened when you increased the concentration of glucose? Why?
 What happened when both beakers started out with the same initial glucose concentration?
Why?
14) Return to the PhysioEx 10.0 main menu.

PHED 2516 – Lab 2: Experiments Worksheet Page 3 of 10

Table 2: Facilitated diffusion results.
Initial Initial Final Final
Diffusion
Concentration Concentration Concentration Concentration
Trial Solutes Carriers Rate
Left Beaker Right Beaker Left Beaker Right Beaker
(mM/sec)
(mM) (mM) (mM) (mM)

1 Glucose 500 2 0 1 1 0.008

2 Glucose 900 2 0 1 1 0.0012
3 Glucose 500 8 0 4 4 0.0023
4 Glucose 900 8 0 4 4 0.0038
5 Glucose 900 8 8 8
8 0

PHED 2516 – Lab 2: Experiments Worksheet Page 4 of 10

Experiment #3 – Osmosis
1) In the PhysioEx 10.0 main menu, click Exercise 1: Cell Transport Mechanisms and Permeability,
then click Activity 3: Simulating Osmotic Pressure. Once the activity opens click on the Experiment
tab. Again, ignore the instructions at the top.
2) On the left side of the simulation, click the + sign beside the Na+/Cl- concentration indicator until it
reads 5mM. Then click the Dispense icon.
3) On the right side of the simulation, click the Deionised Water icon and then the Dispense icon.
4) Click and hold on the 20 MWCO membrane tube to the far right. Drag the tube to the middle of
the beakers at the top of the display and release it.
5) Click the Start icon and observe the concentration levels displayed beside each beaker and the
pressure readings above each beaker.
6) Click Record Data.
7) Record the final concentrations of Na+/Cl- in the left and right beakers (using the numbers beside
the beakers) along with its diffusion rate (from the recorded data) into Table 3 below. Also record
the pressure reading for the left and right beakers (using the numbers above the beakers) into
Table 3.
8) Drag the 20 MWCO membrane back to its original location and click both Flush icons.
9) Repeat for the remaining 3 membranes.
10) Repeat all 4 membranes with 5mM of Albumin.
11) Repeat all 4 membranes with 5mM of Glucose.
12) Repeat the last 200 MWCO membrane and 5mM Glucose trial with 5mM of Albumin in the right
beaker.
13) Answer the following questions as you work through the experiment.
 Did you see an increase in pressure readings for any of the trials? Why would you see an
increase in pressure in the beakers? (Note that the beakers are sealed)
 Did the membrane MWCO have an influence on the pressure readings? Why would the
pressure readings change with different MWCOs?
 Were there any differences observed between the pressure readings of Na+/Cl-, albumin and
glucose? Why was that observation made?
 In the last trial, did your results differ from the other trials? Why would your results differ?
14) Return to the PhysioEx 10.0 main menu.

PHED 2516 – Lab 2: Experiments Worksheet Page 5 of 10

Table 3: Osmosis results.
Final Final Final
Initial Initial Final
MWCO Concentratio Diffusion Pressure Pressure
Concentration Concentration Concentration
Trial Solutes n Right Rate Left Right
(g/mol) Left Beaker Right Beaker Left Beaker
Beaker (mM/sec) Beaker Beaker
(mM) (mM) (mM)
(mM) (mmHg) (mmHg)

1 Na+ / Cl- 20 5 0 5 0 0 170 0

2 Na+ / Cl- 50 5 0 2.5 2.5 0.0093 0 0
3 +
Na / Cl -
100 5 0 2.5 2.5 0.0093 0 0
4 Na+ / Cl- 200 5 0 2.5 2.5 0.0093 0 0
5 Albumin 20 5 0 5 0 0 85 0
6 Albumin 50 5 0 5 0 0 85 0
7 Albumin 100 5 0 5 0 0 85 0
8 Albumin 200 5 0 5 0 0 85 0
9 Glucose 20 5 0 5 0 0 85 0
10 Glucose 50 5 0 5 0 0
0 85
11 Glucose 100 5 0 5 0 0 85 0
12 Glucose 200 5 0 2.5 0.0024 0 0
2.5
Glucose 200 5 0 2.5 0.0024
13 2.5
Albumin 200 0 5 0 5 0 0 85

PHED 2516 – Lab 2: Experiments Worksheet Page 6 of 10

Experiment #4 – Filtration
1) In the PhysioEx 10.0 main menu, click Exercise 1: Cell Transport Mechanisms and Permeability,
then click Activity 4: Simulating Filtration. Once the activity opens click on the Experiment tab.
Again, ignore the instructions at the top.
2) In the middle of the simulation, click the + sign beside each of the Na+/Cl-, Urea, Glucose and
Powdered Charcoal concentration indicators until they all read 5mg/ml. Then click the Dispense
icon.
3) Click and hold on the 20 MWCO membrane tube to the bottom right. Drag the tube to the middle
of the beakers at left of the display and release it.
4) Click the Start icon and observe the concentration levels displayed beside each beaker.
5) Drag the 20 MWCO membrane to the Membrane Residue Analysis area and then click the Start
Analysis icon.
6) Click Record Data.
7) Record the filtrate concentrations of Na+/Cl-, Urea, Glucose and Powdered Charcoal along with
their filtration rates (from the recorded data) into Table 4. Also record whether or not any residue
was present in the membrane (from the recorded data) into Table 4.
8) Click the membrane to return it back to its original location and click the Flush icon.
9) Repeat for the remaining 2 membranes.
10) Repeat all 3 membranes with the pressure set to 100mmHg. (click the + sign beside the pressure
indicator)
11) Answer the following questions as you work through the experiment.
 How did the membranes affect the filtration rate?
 How did the membranes affect which solutes ended up in the filtrate (the solution that
passed through the filter)?
 Where there any differences between the original solute concentrations and those in the
filtrate? Why?
 How did increasing the pressure affect the filtration rate and filtrate concentration?
12) Return to the PhysioEx 10.0 main menu.

PHED 2516 – Lab 2: Experiments Worksheet Page 7 of 10

Table 4: Filtration results.
Initial Filtrate Filtration Residue
MWCO Pressure
Trial Solutes Concentration Concentration Rate Present
(g/mol) (mmHg)
(mg/ml) (mg/ml) (ml/min) (Y/N)

Na+ / Cl- 20 50 Hg 5 0 1 y
Urea 20 50 Hg 5 0 1 y
1
Glucose 20 50 Hg 5 0 1 y
Powered Charcoal 20 50 Hg 5 0 1 y
Na+ / Cl- 50 50 Hg 5 4.81 2.5 y
Urea 50 50 Hg 5 0 2.5 y
2
Glucose 50 50 Hg 5 0 2.5 y
Powered Charcoal 50 50 Hg 5 0 2.5 y
+ -
Na / Cl 200 50 Hg 5 10 y
4.81
Urea 200 50 Hg 5 4.74 10 y
3
Glucose 200 50 Hg 5 4.39 10 y
Powered Charcoal 200 50 Hg 5 0 10 y
+ -
Na / Cl 20 100 Hg 5 0 2 y
Urea 20 100 Hg 5 2 y
4
0
Glucose 20 100 Hg 5 0 2 y
Powered Charcoal 20 100 Hg 5 0 2 y
Na+ / Cl- 50 100 Hg 5
4.81 5 y
Urea 50 100 Hg 5 y
5 0 5
Glucose 50 100 Hg 5 0 5 y
Powered Charcoal 50 100 Hg 5 5 y
0
Na+ / Cl- 200 100 Hg 5 4.81 y
20
Urea 200 100 Hg 5 4.74 20
6
y
Glucose 200 100 Hg 5 4.39 20 y
Powered Charcoal 200 100 Hg 5
0 20 y

PHED 2516 – Lab 2: Experiments Worksheet Page 8 of 10

Experiment #5 – Active Transport
1) In the PhysioEx 10.0 main menu, click Exercise 1: Cell Transport Mechanisms and Permeability,
then click Activity 5: Simulating Active Transport. Once the activity opens click on the Experiment
tab. Again, ignore the instructions at the top.
2) On the lower left side of the simulation, click the + sign beside the Na+/Cl- concentration indicator
until it reads 6mM and click the + sign beside the K+/Cl- s concentration indicator until it reads 4mM.
Then click the Dispense icon. This adds Na+ and K+ to the solution inside of a cell
3) On the lower right side of the simulation, click the + sign beside the Na+/Cl- concentration indicator
until it reads 6mM and click the + sign beside the K+/Cl- s concentration indicator until it reads 4mM.
Then click the Dispense icon. This adds Na+ and K+ to the solution outside of a cell
4) On the cell in the middle of the beaker, click the + sign below the Na+/K- Pump indicator until it
reads 500 units. This creates a membrane with that number of Na+/K- pumps.
5) Click the + sign beside the ATP dispenser at the top of the simulation until it reads 1mM. Then click
the Dispense ATP icon.
6) Click the Start icon and observe the concentration levels displayed beside each beaker.
7) Click Record Data.
8) Record the final concentrations of Na+/Cl- (or Na+) and K+/Cl- (or K+) (using the numbers beside the
cell and beaker) along with their diffusion rates (from the recorded data) into Table 5. Also record
final amount of available ATP (from ATP dispenser) into Table 5.
9) Click the membrane to return it back to its original location and click both Flush icons.
10) Repeat the previous trial of 6mM Na+/Cl- & 4mM K+/Cl- in the cell, 6mM Na+/Cl- & 4mM K+/Cl- in
the beaker, and 1mM ATP with 1000 Na+/K- pumps.
11) Repeat the previous trial of 6mM Na+/Cl- & 4mM K+/Cl- in the cell, 6mM Na+/Cl- & 4mM K+/Cl- in
the beaker, and 1000 Na+/K- pumps with 4mM of ATP.
12) Repeat the previous trial of 1000 Na+/K- pumps and 4mM of ATP with 6mM Na+/Cl- & 10mM K+/Cl-
in the cell, 10mM Na+/Cl- & 4mM K+/Cl- in the beaker.
13) Answer the following questions as you work through the experiment.
 For equal concentrations of Na+/K- inside and outside of the cell, what would you expect
from a simple diffusion stand point? What did you observe?
 How did adding more Na+/K- pumps effect the final Na+ or K+ concentrations and diffusion
rate?
 How did adding more ATP effect the final Na+ or K+ concentrations and diffusion rate?
 In the 1mM ATP trials, why does the transportation of Na+ and K+ stop before all of the Na+
or K+ is remove from their original beaker (note the final ATP concentrations in Table 5)?
 How do you know that the phenomenon demonstrated is not just simple diffusion? (Hint:
think about the last trial).
14) Return to the PhysioEx 10.0 main menu.

PHED 2516 – Lab 2: Experiments Worksheet Page 9 of 10

Table 5: Active transport results.
Initial Initial Final Final
Initial Diffusion
Concentration Concentration Concentration Concentration Final ATP
Trial Solutes Pumps ATP Rate
Cell Beaker Cell Beaker Available
Available (mM/sec)
(mM) (mM) (mM) (mM)

Na+ 500 6 6 2.731 9.269 0.0309

1 1 0
K- 500 4 4 0.0206
6.179 1.821
Na +
1000 6 6
6 9.864 0.0548
2 1 0
K- 1000 4 4
6.576 1.424 0.0365
+
Na 1000 6 6 0
3 4 12 1.001 0.0074
K -
1000 4 4
8 0 0.0049
Na+ 1000 6 10 0 16 0.0099
4 4 1.001
-
K 1000 10 4 14 0

PHED 2516 – Lab 2: Experiments Worksheet Page 10 of 10