Leishman stain: Principle, Procedure, use

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: Principle, Procedure, use
Leishman stain

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.

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