OBJECTIVES
- To prepare temporary slide.
- To Show Stomata.
- To Show cells of ventral peel of leaf.
MATERIALS REQUIRED
Fresh leaf of Senna tora (or Bidens pilosa /Tradescantia /Celosia cristata/ Dioscorea bulbifera) or other soft dicot leaf, Safranin, Dropper, Glycerol, No. 0 painting round tip brush, forceps, needle, blade, scissor, wash bottle, petri dish, watch glass, cover glass, slide, compound microscope .
THEORY
Stomata are specialized structures found on the ventral surface of leaves and stems of plants. They play a crucial role in gas exchange, allowing the intake of carbon dioxide (CO2) for photosynthesis and the release of oxygen (O2) and water vapor (H2O) through transpiration. The study of stomata can provide valuable insights into plant physiology and adaptation to various environmental conditions.
The experiment aims to prepare a temporary slide of stomata from the ventral peel of a dicot leaf. The ventral side of the leaf refers to the lower surface, which is usually more suitable for stomatal observation due to higher stomatal density
PROCEDURE
- Obtain a fresh dicot leaf (Senna tora), that is free from damage or disease.
- Carefully remove a small section of the ventral peel of the leaf using forceps. Ensure that the peel contains intact stomata.
- Cut the ventral peel in square about 3-4 mm length by scissor then keep in watch glass containing distilled water.
- Then transfer peel through brush to another watch glass containing 2-3 drops of Safranin and wait for 30 to 60 seconds.
- Then wash stain by transferring tissue to another watch glass containing clean distilled water for 2 or 3 times to remove stain completely.
- Take a clean microscope slide and pour a drop of glycerin at the center of slide.
- Then transfer tissue to slide containing glycerine.
- Then apply cover slip with needle to prevent bubble.
- Then watch under the compound microscope with 10X objective at first then at 45X objective.
- You find many stomata with guard cell, then draw diagram in practical note.
OBSERVATION
Many stomata seen with kidney shaped guard cell in epidermis.
RESULT
Most of the stomata found in ventral surface of leaf.
CONCLUSION
Stomata present in ventral surface of leaf successfully observed by making temporary slide.
PRECAUTIONS
- Handle the fresh dicot leaf with care to avoid damage or tearing.
- Ensure the microscope slide and coverslip are free from dust or debris.
- Take care not to introduce air bubbles between the peel and coverslip when mounting the slide.
- Don’t stain more than 1 minute to prevent over staining
- Keep leaf and peel in water to prevent dehydration.