OSMOSIS IN CELLS OF AN ONION & USING STAINS

Cells are so tiny that a microscope is needed to see them. Often it is necessary to use the High
Power (HP) objective lens. Cells are usually transparent and sometimes a stain has to be
added to the cells, which reacts with chemicals inside the cells, changing their colour, and
making them visible.

1. DECIDING WHICH STAIN TO USE

Not all stains work well in all tissues, so our first decision is to decide which stain we should
use, either methylene blue or iodine. For the next investigation, we specifically want to see the
central vacuoles of the onion cells.

Collect the following materials and equipment and take them to your work-place:

Materials Equipment Chemicals

piece of red onion leaf microscope methylene blue stain
plastic pipette iodine stain
slide(s) distilled water
cover slip(s) 5% NaCl (Salt) solution
needle or scissors

What you do, to prepare 3 slides for comparison

1. Carefully read and follow these instructions:
• Collect a small piece of onion and identify which is the inside of the leaf
• With a knife or sharp point, lift up a piece of the inner layer of the leaf. (This is called
the inner epidermis). This layer is just one cell thick.
• Cut a square of the epidermis, about 2mm x 2mm – much smaller than you think!
• Lay this square onto the middle of a clean slide. Check that the layer of cells do not
roll over.
• Add 2 or 3 drops of distilled water to the epidermis.
• Put a cover slip on top. Make sure that you do this properly. Remember how?, You do
not want bubbles to appear.
• Examine the slide with Low Power and then Medium Power. Maybe, carefully, go to
High Power.

2. You can use stains to make the inside of a cell more visible. The best two stains are
iodine and methylene blue.

3. Prepare another piece of onion ‘skin’ and add two drops of methylene blue instead of
water. Leave the stain for 3 minutes before putting on the cover slip. Examine the cells
with the microscope.

4. Repeat this a third time, using iodine stain.

(You can share this work on your table but make sure that the 3 investigations are a fair test
by following the same procedures.)

Show your 3 slides to John [5 points]

Turn over
Year 9 Science Cells and Microscopes
Evaluating the results
• How will you decide which stain works best? Use a 5 point scale like the one below:
1 2 3 4 5
vacuoles still cells show up
difficult to see very clearly

• Put the results into this results table:

Stain used Score on scale 1 - 5

Water
Iodine
Methylene blue

[2 points]
• When you compare two different variables in an investigation, you must make sure that
your investigation is a FAIR TEST. Here you were comparing the two stains, iodine and
methylene blue. Was your investigation a fair test? If you answer “Yes!” then write why.
If you answer “No!” then write why.

Was the investigation a FAIR TEST? YES or NO. ........................................................................

Explain why you have answered “Yes!” or “No!” .........................................................................

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[3 points]

2. OSMOSIS – a form of diffusion

With your group, decide which stain will make it easiest for you to see the individual onion
cells, including the central vacuole.

Membranes have an important function in all cells, to control what passes into and out,
through the membranes. Not all chemicals and particles can pass through all membranes.
There are 3 main ways in which substances can pass through membranes:
1. If there is a concentration gradient, with a higher concentration of the substance on
one side than the other, then this substance may be able to simply flow through the
membrane, with the membrane unable to control the happening. This is called
diffusion. Very few substances can pass through a membrane in this way.
2. Some chemicals which have a concentration gradient but cannot simply diffuse across a
membrane may be helped through the membrane by special proteins. This is like a flip-
flop system! No energy is used by the cell. It is called facilitated diffusion.
3. Sometimes a cell needs to move chemicals or substances where there is no concentration
gradient. Energy is required in this case to actively bring substances across the
membrane, through special protein channels. This is called active transport.

Year 9 Science Cells and Microscopes

 4. Many cells have the capacity to engulf liquids and solid substances, using their
membranes. Think about those phage bacteria which are able to engulf viruses. This is
called bulk transport.

And then we have OSMOSIS!

Osmosis is actually very simple to understand. Osmosis is the movement of only water from a
solution on one side of a membrane to the other side. If there is more water on one side than the
other, then water will diffuse across the membrane. How can there be more water on one side
than on another? We need to know about solutions. (Cue – powerpoint! And a YouTube video:
https://www.youtube.com/watch?v=L-osEc07vMs
Cornell notes on membrane transport, solutions & osmosis. 1 grade in Idukay. [10 points]

Investigation into OSMOSIS

1. Prepare a slide of a piece of red onion epidermis, exactly as you did previously, using the
stain that gave you the best results. Place a cover slip on the slide.
2. Use a microscope to check that you can properly see the cells.
3. Carefully remove the slide from the microscope and place it on a flat surface.
4. Use a pipette to place 2 drops of 5% NaCl solution along one side of the cover slip – in
contact with the liquid beneath the cover slip but not on the coverslip.
5. Now place a piece of paper towel along the other side of the coverslip. This will draw the
5%NaCl solution through the slide, so that the onion cells are flooded by salt solution.

5% NaCl solution

6. Carefully replace the slide on the microscope and examine the cells. What do you see?
7. Describe what you see and make a drawing of a small group of cells in the space below.

Describe what you see: ………………………………………………………………………………………………………..

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[2 points]

Labelled drawing

[3 points]
Turn over
Year 9 Science Cells and Microscopes
 8. Remove the slide from the microscope.
9. Lift off the piece of paper towel and dry the slide on both sides of the coverslip. (Be careful not to
move the coverslip and the onion cells.)
10. Now use the pipette to place drops of distilled water along one side of the coverslip and draw the
distilled water through the piece of onion with a paper towel, as you did before.
11. Describe what you see and make a drawing of a small group of cells in the space below.

Describe what you see: ………………………………………………………………………………………………………..

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[2 points]

Labelled drawing

[3 points]

Conclusion – the hard part!

If this investigation has worked out properly for you, you should have been able to observe
OSMOSIS in action. Write a conclusion – a summary of your observations – using the words
OSMOSIS, TURGOR PRESSURE and PLASMOLYSIS to describe what you have seen in
this investigation.

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[5 points]
[Total 25 points]
Year 9 Science Cells and Microscopes