Leishman stain principle and procedure

Leishman stain is a commonly used stain in the hematology lab. It is used to identify the white blood cells, malaria parasite, cutaneous leishmania, and Trypanosoma species.

Leishman stain preparation Requirements

Following are the requirements of Leishman stain preparation.

1. Methanol: Acetone free, 100 ml
2. Leishman stain: High purity powder 0.2 gram.
3. Conical flask: Used for mixing methanol and stain.
4. Funnel
5. Mortar and pestle: used for the grinding the powder.

Leishman stain procedure

• First of all, weigh 0.2 g of Leishman powder stain and transfer it to mortar for grinding.
• Grind, stain with 20 ml methanol, and allow it to settle. Transport the supernatant through a filter paper to the flask.
• Add another 25 ml alcohol to the mortar, containing residual stain. Repeat the grinding procedure. Allow it to settle and transfer it to the flask.
• Note: Repeat the procedure until all of the methanol has been used and most of the stain dissolved.
• Place the flask in the water bath at 50 C for 25 minutes.
• Filter the stain into a borosilicate glass bottle for RIPENING.
• Leave it for 2 to 3 days in the dark at room temperature.