The procedure of preparing Leishman stain(stock solution) – procedure, storage, labeling

Equipment and chemicals required

  • Leishman stain powder
  • Absolute methanol
  • Spatula
  • Graduated cylinder
  • Glass or plastic funnel
  • Screw – capped, dark or amber glass bottle(250ml)
  • Analytical balance/weighing scale
  • Mortar and pestle
  • Shaker


  • Weigh 0.15g of Leishman stain power on an analytical balance or weighing scale
  • measure 100ml of methanol into a measuring cylinder
  • place powder in motor, grind with methanol until a smooth suspension is observed. This should be done slowly and take precautions not to mix with water
  • put it into the dark or amber-colored glass bottle through a funnel
  • Gently pour about 20ml of methanol through the same funnel, ensuring that all the stain is washed into the bottle
  • Tighten the screw-cap on the bottle and shake it in a circular motion for 2-3 minutes(or use a shaker)to dissolve the stain crystals in methanol solution
  • Warm the content at 50 Celsius in a water bath for 15minutes with intermittent shaking
  • Add the remaining amount of methanol to the mixture through the funnel and make the volume 100ml
  • Tighten the screw – cap on the bottle and continue shaking for few minutes
  • stain is ready to use after one week
  • Filter the stain using filter papers before use


  • Dark bottle
  • Tighten the screw–cap on the bottle to prevent absorption of water vapor from the air, and store it in a cool place away from sunlight. Protect from light. Store at 2-2.25 Celcius appropriated stored solution can be used about 3 months


  • Label the bottle clearly. label should include name of the chemical(Leishman stain stock solution)
  • Name of the person who prepared it
  • date of preparation
  • date of expiry



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