Blood Smear Preparation and Staining Practical Lab

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Total WBC Count: https://www.youtube.com/watch?v=xyhbIPSLBsA&t=364s
Total RBC Count: https://www.youtube.com/watch?v=0f9p9JX4qJk&t=130s
DLC: https://www.youtube.com/watch?v=VFKm_kMTf50
Haemocytometer Use: https://www.youtube.com/watch?v=7IpE33WPRrc&t=43s
Haemometer Hb estimation: https://www.youtube.com/watch?v=6CqptdZyUaU&t=45s
Blood Smear Prep and Staining: https://www.youtube.com/watch?v=KSs0SMfERuA&t=38s
RBCs in iso, hypo and hypertonic solns: https://www.youtube.com/watch?v=urYJAPGUpuw
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Thin Peripheral Blood smear preparation and Staining
Blood is a type of fluid connective tissue that carries nutrients and oxygen to various cells in the body and at the same time transports metabolic waste away from the same cells.
Vertebrate blood is composed of blood cells suspended in plasma.
Blood cells mainly consists of Red Blood cells or Erythrocytes or RBCs, White Blood cells or Leucocytes or WBS and and Platelet cells. RBCs are the most abundant blood cells. They impart a red colour to the blood.
In this experiment, we will be covering the procedure on how to prepare a thin blood smear and stain it with an appropriate stain for the morphological and quantitative study of the various cellular elements of blood.
For this experiment, we will need the following:
Blood sample
Glass slides
Leishman’s stain or Wright’s Blood stain or Giemsa’s stain
Distilled water
Droppers
Petridish or Coplin jar
Compound light microscope
Immersion oil
Cotton and rubbing alcohol
Blood lancet or a pricking device

Take two glass slides and clean it with 90% alcohol
Disinfect a finger with alcohol and prick with a sterile lancet or a pricking device
Place a tiny droplet of the blood at the centre of one corner of one of the glass slides.
Using the second pre cleaned slide, touch the drop of blood while inclining the slide at a 45 degree angle.
Now gently and briskly move the inclined slide towards the other end of the slide containing the blood drop in order to obtain a smear.
A properly smeared blood appears roughly tongue shaped as seen here.
Allow the smear to air dry for a minute or so.
Place the slide in a petri dish with the blood smear facing up.
Add several drops of the Leishman’s stain to the blood smear and cover the petri dish.
Alternatively, you may fill a coplin jar with Leishman stain and immerse the slide in the stain.
Stain for 1 minute and then add twice the volume of distilled water to the stain.
Allow the water and the stain to properly mix and keep it aside for about 10minutes.
Drain the slide and wash with distilled water.
Proper staining results in a blood smear that is rose pink in colour.
Air dry the slide while keeping it in an inclined position.
The slide is now ready for observation under the microscope.
Observe and study the various blood cells under 40x and 100x objective.
For 100x , place a drop of immersion oil on the slide, gently lower the 100 x objective until it touches the oil surface. Hereon, use the fine adjustment knob for focussing.

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