Preparation of stomatal slide of leaf to study stomata and epidermis

Experiment to Observe Temporary Mount of a Leaf Peel to Show Stomata and epidermis
 
Experiment:

Objective:
To prepare a temporary mount of a leaf peel to show stomata.

Apparatus and Materials Required:

A plant or specimen, forceps, needles, watch glasses, Petri plates, glass slides, a dropper,
coverslips, a brush, blotting paper, safranin, glycerin and a compound microscope.

Theory:
Stomata are small openings found widely scattered on the epidermis of leaves and young stems.
They are mostly found on the lower surface of a dicot leaf and on both the surfaces of a monocot
leaf. Stomata regulate the exchange of gases and water vapor between the atmosphere and
leaves.

Procedure:
1. Remove a healthy leaf from the plant.
2. The leaf peel of lower epidermis is taken and cut with scissors or blade into pieces. These
pieces transferred with the help of paint brush into petri dish containing water (do not
keep peel without water they will dry)
3. Remove a part of the peel from the lower surface of the leaf. You can do this by folding
the leaf over and gently pulling the peel apart using forceps. Keeps the peel in a watch
glass containing water.
4. Put a few drops of safranin stain in a watch glass
5. After 2-3 minutes take out the peel
6. A piece of leaf peel is transferred into another dilute solution of safranin
7. Place peal into clear water to remove excess water the remove peel from water
8. Place it on a clean glass slide.
9. Put a drop of glycerin over the peel and place a clean coverslip gently over it with the
help of a needle.
10. Remove the excess stain and glycerin with the help of blotting paper.
11. For inner parts study leaf peel is observed under high power (45x) of microscope.
12. Observe image take photograph and note readings regarding setting of microscope.

Precautions
1. The epidermal peel should be taken from freshly plucked leaf
2. Take the epidermal layer from the lower side of leaf it contains more stomata
3. Always use clear glass slide

Limitations of peel method: ‐

1. Not all leaves are easy to peel
2. Frequently mesophyll cells come off with the epidermal layer, giving more than one cell
layer and making identification of the epidermal cells difficult and confusing. 

Figure 1: Preparation of slide of stomata

Figure: 2 Lower epidermis structure

Figure :3 Visual representation of microscopic view of epidermis from microscope