practical write up smear

1. Tansen School of Health Science
Practical write up
Subject: Haematology
Title: blood smear and staining
Aim: to prepare the smear and stain by Leishman and wright
stain
Introduction:
Bloodsmear:- a blood smear is a blood test used to look for abnormalities in blood
cells. The main blood cells that the test focus on are:-
Red cells,white cells,platelets.
This test provides information on the shape and size differential count platelets
count ,which can help doctors diagnosecertain blood disorders or other medical
condition.
Staining:- by the help of staining the laboratory person can identify the blood cells.
Materials required:
Slides
Speader
Blood with mixed EDTA
Dropper
Test tube rack
Something to clean
Staining rack
Reagents:
Leishman stain
Wright stain
Buffer solution

2. Procedure:
1) Take a clean slide
2) Place a drop of well mixed blood 1-2cm from one end of the slide in the centre
3) Choose smooth slide as a spreader, place this slide near the drop of blood at an angle of 30-
45O
4) Move spreader back to make contact with the drop of blood
5) Move the spreader forward in a smooth and rapid manner to obtain a nice regular smear.
Size of the smear should around 3/4th of the slide
6) Allow to dry
For Leishman’s stain:-
1) Prepare smear and air dry it.
2) Cover the smear with stain and wait for 2 mints for fixation.
3) Dilute with double volume of buffer water and wait for 7-10 minutes for staining.
4) Wash the smear with tap water.
5) Clean the back side of slide and then drain and air dry the smear
6) Examine under microscope.
For wright’s stain:-
1) Prepare smear and air dry it.
2) Cover the smear with wright’s stain for 1-2 minutes (fixation time), taking care that it should
not dry on the slide. Main aim is fixation.
3) Dilute with an equal volume of buffer water, PH 6.8.
4) Keep the diluted stain for 3-5 minute(staining time)
5) Wash the smear with buffer water or tape water.
6) Clean the back side of slide by using tissue paper.
7) See under microscope for proper staining i.e. to check the smear under or over stained.
8) Let it dry at RT by keeping upright position in slide rack for drainage and examine under
microscope.

3. Internal Quality Control:
1) Smear should be tongue shape.
2) Slide should be clean and dry before use.
3) Blood should not over flow.
4) Clotting blood shouldn’t be used.
5) Solution shouldn’t kept on refrigerator.
6) Stain manufactured date should be checked. etc
References:
Cheesbrough, M., 2006. District laboratory practice in tropical countries. Cambridge university press.
Godkar, P.B. and Godkar, D.P., 2006. Textbook of medical laboratory technology. Bhalani publishing house.
Dacie and Lewis, Practical Haematology.