Practical No 3

PLASMOLYSIS AND DE-PLASMOLYSIS IN BLOOD.

PREPARATION OF BLOOD SMEARS.
 Plasmolysis
´Plasmolysis is defined as the process of contraction or shrinkage of the
protoplasm of a blood cell and is caused due to the loss of water in the cell.
The word Plasmolysis was generally derived from a Latin and Greek word
plasma – The mould and lusis meaning loosening.
 Deplasmolysis
Deplasmolysis is the process that occurs when a plasmolyzed cell (a cell
that has undergone plasmolysis) is placed in a hypotonic solution, leading to
the absorption of water back into the cell. This results in the cell regaining
its turgid state.
Materials Slides,
Ringer solution,
3% NaCl solution (Hypertonic solution),
Distilled water (Hypotonic solution),
Dropper
Microscope.
 1.Take three slides and mark these A, B, and C.

Procedure 2.Cleanse the tip of your ring finger with spirit.

3.Prick your fingertip to get a drop of blood.
4.Transfer this blood into about 0.5ml-1ml Ringer
solution.
5.With the help of a dropper, put one drop each
into slides A, B, and C.
6.Add 3% NaCl solution to slide B.
7.Add distilled water to slide C.
 1. In Slide A, you will observe that the RBCs
retain their biconcave shape and size because

Observations the ringer solution is isotonic so there is no

osmosis.
2. In slide B, you will observe, that the cells
shrink due to exosmosis (of water), a process
called plasmolysis.
3. In slide C, you will observe, that the cell swells
up, and increases in size a process called DE
plasmolysis.
Preparation of Blood Smear
 1. Cleanse the tip of your ring finger with spirit.
2. Prick your fingertip to get a drop of blood.

Procedure
3. Touch the next drop of blood with a clean slide. If blood does
not well up, gently squeeze the finger.
4. Bring a clean spreader slide, held at a 45° angle, toward the drop
of blood on the specimen slide.
5. Wait until the blood spreads along the entire width of the
spreader slide.
6. While holding the spreader slide at the same angle, push it
forward rapidly and smoothly.
7. Wait until the film is completely dry before staining. Fix the thin
film with methanol (100% or absolute) and let it dry completely
before staining.
 Observations
 In the prepared blood smear slide we will

Observations observe:
 Erythrocytes: biconcave disc-shaped.
 Platelets: small discoid-shaped 1.5 -3 µm.
 Monocytes: large irregularly shaped and folded.
 Lymphocytes: can look like monocytes but are
usually smaller and do not have a kidney-bean-
shaped nucleus.
 Neutrophils: bigger than RBCs, multilobed (2-5
lobed) nucleus