Instructions - Osmosis & Diffusion Experiment

Aim:
To give the student a better understanding of the workings of diffusion and osmosis by performing several
experiments. Also to give the student experience in the use and handling of common lab equipment.

Background Information:
Diffusion is the process where solute particles spontaneously spread out to even their distribution. The
particles naturally flow, through kinetic action, from areas oh high concentration to low concentration. A small
amount of sugar dumped in one end of a pool will spread out until the concentration throughout the entire
pool is equal. Osmosis is a special case of diffusion where the solute is prevented from spreading out to
equalize concentrations, but the solvent shifts instead. Osmosis is a physical process in which a solvent,
normally water, moves across a semi permeable membrane separating two solutions of different
concentrations. The water will move from the region of higher water potential (lowest concentration of solute)
to a region of lower water potential (highest concentration of solute). Semi permeable refers to a membrane
through which water can travel but not the solute, such as sucrose. Plasmolysis occurs when a plant cells
membrane shrinks away from its cell wall. This phenomenon occurs when water is drawn out of the cell and
into the extracellular (outside cell) fluid. The Water Potential is the potential energy of a solution relative to
pure water. The water potential of pure water is considered to be zero and all solutions will have a negative
value. Thus water will flow from a higher water potential to a lower water potential (from 0 to any -value).
Water potential is the sum of A) pressure potential (physical pressure) and B) solute potential.

water potential = pressure potential + solute potential

The pressure potential is simply the atmospheric pressure measured in bars. The solute potential is
calculated by using the equation:

solute potential = -iCRT

Where i = the ionization constant, C = the molar concentration in mol/L, R = the constant 0.0831 L bar/mol K and T =
temperature in K. The water potential for a 1.0 M solution of sucrose at 1 atmosphere pressure and a temperature of 25
C would be

water potential = pressure potential + solute potential

water potential = 0 bars + (-1 x 1.0 mol/L x 0.0831 L bar/mol K x 298 K)

-24.8 bars

The pressure potential is zero at 1 atmosphere. A material such as sucrose does not ionize thus the ionization constant
is 1 (NaCl breaks up so it would be 2, CaCl2 would be 3, etc.).

In this lab you will determine the water potential of potato cubes and observe plasmolysis in an Elodea plant.

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Many different solutions are used in this lab. This is a

diagram of 6 beakers holding solutions from distilled
water to 1.0 M in sucrose. The potato cubes will be
placed in these solutions and the gain or loose of mass
will be noted
---*---
 Plastic wrap will be
used to cover the
beakers while the potato
cubes adjust to their
solutions.
---*---
An electric balance
will perform all
massing during this
lab.

---*---

This diagram displays the tongs used to

transfer the potato cubes, the drying bowl
used to remove exterior dampness and the
bowl which holds the potato samples.

---*---

A microscope will be used in the

second part of this lab where
plasmolysis is observed. A close-
up view is provided as seen
through the microscope.

---*---

To promote
plasmolysis,
a salt water
solution is
used. A
cotton swab
is used to
draw the
solution
under the
cover slip
on a slide.
The water is
used to
 reverse this
process.
-

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If you get into trouble and perform some procedure that causes the lab to fail(lab equipment will no longer
operate), you can press the "Reset" button and the simulation will return to the starting position.

---*---

PROCEDURE:

ROOM 1: Water Potential

1) You can adjust the background shading by clicking on the "Special" button to the right and selecting "Background".
Click on the "Special" button and select "Print Blank Report" to obtain a web page that can be printed and used as a
lab report. (the program will not be interrupted)

2) Wear your goggles. Remove the plastic wrap from the bowl of potato cubes. This cover has prevented the cubes from
drying out after they were prepared. Pick up the tongs and hold the clamping end over a group of potato cubes in the
bowl. Press "p" and the tongs should grab a group of 4 cubes.(this may take a little practice) Carry the tongs, holding
the potato cubes, over to the "Drying Cotton" bowl and press "p" to drop them in the bowl. This will remove any outside
moisture on the cubes. Pick up the cubes and carry them over to the electronic scale and drop them on the massing
pan. Record their initial mass in the 0.0 M (distilled water) blank. Pick up the cubes and drop them in the 0.0 M beaker.

Note: since the cubes are not marked, perform the above steps completely before picking up another potato sample!

3) Perform procedure #1 for the other 5 potato samples, dropping each in a different beaker. Record their initial masses.

4) Pick up a new piece of plastic wrap and cover beaker 0.0 M. Do the same for the other 5 samples. This plastic wrap
will prevent evaporation during the long soaking period.

5) Click "Run" on the timer and the 24 hour soaking period will begin. Note: virtual time can pass by at a much faster
rate than real time.

6) When the soaking period is over, remove the plastic wrap from beaker 0.0 M. Pick up the cubes, dry them, drop them
on the electronic balance and record their final mass. Repeat this for the other samples (you can dump the cubes back
into their original bowl).

This lab takes place at 1 atmosphere pressure and a temperature of 25 °C.

You have completed the first half of the experiment. Press the "Next Room" button to move on to the next half.

ROOM 2: Plasmolysis
A slide of Elodea has been prepared and placed on the microscope stage. The Elodea tissue was mounted on the slide
using normal pond water. Turn on the microscope light by clicking on the front switch. The large circle represents the
view from the microscope. The small rectangle to the circles lower right is a real time cell that will show plasmolysis in
action.(the circle will just show the results)

7) Draw a picture of several of the complete cells seen in the microscope view.

8) Pick up the eyedropper of salt water and place the tip on the right side of the slide, at the right junction where the
coverslip and slide end.
Press "p" to place a drop in this region. You can tell if you miss! The salt water will not automatically be drawn under the coverslip.
Pick up the cotton swab and place the lower right cotton end on the opposite side of the coverslip. This will draw the salt water
under the coverslip.

Note: observe the change that the "Real Time" cell goes through.

9) Draw a picture of several of the complete cells seen in the microscope view.

10) Repeat this procedure but use the water eyedropper.

11) Record and calculate the values asked for on the lab sheet and any handouts given by your instructor.