Leishman's staining

LEISHMAN'S STAINING TECHNIQUE

  • Dry the thin blood smear in air.
  • Cover the smear with a known quantity of the stain (normally, 8-10 drops) and allow to remain for 1 minute.
  • Add double the quantity of neutral distilled water and mix it well with the stain by means of blowing air through a pipette and allow to remain for 10-15 minutes.
  • Wash off the stain quickly and allow the slide to dry in a vertical position and then examine under oil immersion objective.

Staining techniques

  • Leishman staining
  • Stain preparation

Leishman’s stain powder - 150mg

  • Absolute methonol (acetone free) - 100 ml
  • Grind the Leishman stain powder with a little methanol slowly in a pestle and mortar and then add full quantity of methanol and filter it. Ripening period is about 3-4 weeks. Store it in amber colored bottle.

Leishman and Giemsa Cocktail stain

  • Leishman’s stain powder - 150 mg
  • Giemsa stain powder - 30 mg
  • Methanol - 100 ml
  • The stain solution is prepared as described above.

Staining procedure

  • Air dried film is placed on the staining rack and flooded with Leishman stain
  • Allow it to act for 30 seconds to one minute for fixation
  • The stain is diluted nearly by twice of its original volume with distilled water or buffer.The pH of the buffer should be 7.2 for mammalian smear and 6.8 for avian blood
  • Blow the solution until it forms an uniform mixture
  • Allow it to act for 20 minutes
  • Wash carefully with tap water dry it and examine first at low power and then at oil immersion, especially at the fringed edge and border
Last modified: Monday, 4 June 2012, 5:42 AM