This is a scan of maple leaf vascular tissue, done with a normal flat-bed scanner.

Method: Preparing the leaf was the difficult and time-consuming part. The leaf was boiled for several hours until the cells started to separate. I then carefully lifted the leaf out of the pot and placed it on a plate with water. The soft tissue was then removed with a stiff brush, trying not to damage the delicate veins. The veins were then rinsed in alcohol to remove the remaining chlorophyll, washed in water to remove the alcohol. The alcohol also shrinks the structures, but it will expand again when washed in water. The leaf skeleton was then, pressed and dried. Not all leaves work equally well! The leaves of some plant species are so stiff that the cells do not want to come off when boiled. Don’t waste your time on these leaves.

A confession: Because the stem of the leaves come off very easily, I had to scan it separately and then integrate it into the picture later using some photo editing. I could not scan the veins and the stem at the same time, because it then would not be flat on the scanner. You will also notice that some parts of the leaf are not in focus. This too is because the leaf was not completely flat on the scanner.

For teachers and parents: Boil some leaves with your students/children and let them prepare the leaf skeleton. Then observe the leaf skeleton under the stereo microscope.

Maple leaf veins after the removal of the soft tissue. The leaf was dried and then scanned at high resolution.

This picture shows the tip of a maple leaf. Note that not all leaves can be processed this way.

This is a simple but somewhat time-consuming preparatory technique. It is possible to isolate the vascular bundles of certain leaves and prepare them for microscopic observation. The prepared leaf veins make an ideal specimen for stereo microscopy. The microscope allows the students to perform a quality-check of their preparation. You may be interested in the “Virtual Microscope”, which allows you to zoom into the leaf veins: Virtual microscope: maple leaf skeleton

Materials: Maple leaves, hot plate, cooking pot, eating plates, small but stiff brush or toothbrush

Method:

1. Let the leaves simmer for 1-2 hours. Periodically check the leaves by carefully rubbing them between your fingers. They should start to feel slimy and you should be able to rub off some of the surface plant tissue.
2. Carefully lift out the leaves. They are now very delicate and they tear easily. Put one leaf on one dish each.
3. Add a bit of water to the leaf on the dish. Use the brush to carfully remove the soft plant tissue of the leaf. The brush presses the leaf against the plate. This gives the leaf stability. Use the fingers of the other hand to prevent the leaf from moving while brushing. The leaf veins start to appear. Carefully turn the leaf around and remove the plant tissue on the other side as well. The water of the dish starts to accumulate plant tissue and should be exchanged periodically.
4. You now have a delicate network of leaf veins on the plate. Lift it out and place it flat on tissue paper to remove most of the liquid. Press the leaf veins between layers of tissue paper and a book. Otherwise there is the danger that the leaf will warp during the drying process.
5. Observe the leaf veins using a stereo microscope. They can also be observed using a compound microscope using a low magnification. Alternatively it is possible to scan the leaf veins with a flat-bed scanner.
6. Make a quality check. Observe any soft leaf material that has not been removed. Observe any tears and breaks in the leaf veins that were caused by brushing too forcefully.

Alternative method:

• Press the leaf between two books.
• Place the leaved into a solution of washing soda (pH 11 – don’t let children do this!) until they become pulpy and the soft material starts to come off.
• Rinse the leaves and brush off the soft material with a soft brush.

The Efficient Method: Do an Internet search for “skeleton leaves” and buy some ready made ones… Other Ideas:

• Students may also attempt to remove the soft tissue directly under the stereo microscope. In this case the leaf should be placed in a petri dish.
• The cleaned leaf veins can be brightened by washing them in pure alcohol. This removes remains of the chlorophyll. The alcohol also removes water and the network of veins will shrink. Wash the veins in pure water after the alcohol treatment to restore the original size.
• The network of veins can be scanned using a flatbed scanner using high resolution. This also visualizes small structures. A dark background gives a nice contrast.

Troubleshooting: Question: It is not possible to remove the soft tissue of the leaf. Answer: Some leaves can be boiled for hours and still not macerate. Oak leaves are completely unsuitable for this preparatory technique. Try out a variety of different leafs. Alternatively, the leaf may not have been boiled long enough.