A Study of Plasmolysis and Osmosis using Leaves of the Aquatic

Plant Rotala Wallichii

Zia Dostmohamed

219616572

Section A 08

TA: Riddhi Jani

Materials and Methods
The study conducted was the measure the effect of hypertonic solutions on leaf cells of

the aquatic plant Rotala Wallichii. The hypertonic solution CaCl2 was used in this particular

experiment in differing concentrations to try and identify at what concentration plasmolysis

would no longer occur.

A compound microscope was used to analyze the submerged leaf cells and view

occurring plasmolysis. The microscope was set up using Kohler Illumination which provided

even lighting of the specimen, minimized glare, and increased contrast and resolution. The

microscope was handled properly and utilized correctly. Lens paper was used to clean the lenses

to prevent scratching, and when beginning to work with the microscope all power sets were at

their lowest. To preform Kohler Illumination the lens was set to the lowest objective (4X) and

the light intensity dial was set to its lowest setting. The condenser was moved using the

condenser focusing knob to a few millimetres below the stage top and a specimen slide with a

black line was placed and secured on the stage using the stage slide clamps. The stage motion

knob was used to centre the specimen over the opening in the stage, so the light went through it

and it was visible through the lens. The slide should never touch the objective lenses so the

distance between the stage and lenses should always be observed very carefully. The stage was

raised up as far as possible using the coarse focus knobs and then the image was focused and

corrected using the fine adjustment knobs. Next the second objective lens, 10X was used to view

he specimen. The fine focus adjustment knob was used to bring the image into a sharp focus.

This was the final step of setting up the microscope.

Following setting up the microscope, we had to make a slide of the aquatic plant

specimen in hypotonic solution (normal pond water) and compare it with the leaf cells in
hypertonic solution (CaCl2). Using scissors and tweezers, two leaves were cut 1cm in length and

placed on the glass slide 2cm apart. A Pipette was used to put a drop of pond water to the leaf on

the left side and a drop of hypertonic solution (CaCl2) to the leaf on the right side making sure

the two drops were separate and recorded the time the leaves were placed in solution. Each leaf

was viewed with 10x and 40X magnification lens, the cells were to be witnessed under osmotic

conditions. Sketches of observations were made to view the difference between osmotic

condition vs. plasmolysis.

Finally, determinations of thresholds for plasmolysis had to be discovered using

experimentation. Dilutions of stock solution with pond water at different concentrations was

made which was then put on leaves of Rotala Wallichii using a Pipette. The control of the

experiment was the specimen placed in only pond water which showed no visible sign of

plasmolysis occurring, in that the chloroplasts were not grouped. Each slide was left to sit in the

solution for 5 minutes before viewing in order to allow for an equal amount of absorption to have

occurred. 5 specimen slides were created, including the control slide to view plasmolysis at

different concentrations. The concentrations included in this experiment were 2M, 1.5M, 1M,

0.5M, 0M. These concentrations were found using the C1V1 = C2V2 formula and crated using a

mix of CaCl2 and pond water. After producing four calculated dilutions, each leaf cell was

studied for plasmolysis and viewed to see which concentration had the no effect on cells to

undergo plasmolysis.

After conducting the experiment, the microscope was turned off and placed back to its

starting state, all the equipment was rinsed and put back on the tray. The leaves of Rotala

Wallichii were disposed of properly in the gargbge.

Results
Table 1. Leaf cells submerged in hypertonic solutions of the same total volume and different concentrations to view
if plasmolysis occurred

CaCl2 Stock Working Volume of Volume Volume of Did

Concentration Solution Working of CaCl2 Water Plasmolysis

Concentration Solution Occur

2.0M 2.0M 5 mL 5 mL 0 mL Yes

2.0M 1.5M 5mL 3.75 mL 1.25 mL Yes

2.0M 1.0M 5 mL 2.5 mL 2.5 mL Yes

2.0M 0.5M 5 mL 1.25 mL 3.75 mL Yes

2.0M 0.0M 5 mL 0.0 mL 5.0 mL No

Conclusion
We determined that CaCl2 at a concentration of 0.5M triggers plasmolysis in Rotala Wallichii leaves,
where plasmolysis was measured as occurring.