Giemsa's staining

GIEMSA'S STAINING TECHNIQUE

Giemsa stain

Stain preparation

  •  Giemsa powder - 1 G
  •  Glycerol (AR) - 66 ml
  •  Methanol (AR) - 66 ml
  •  Azur II - 0.2 G
  •  Take 54 ml of glycerol and place in a clean round bottom flask containing clean glass beads
  •  Add 1.0 gram of Giemsa powder
  •  Heat to 60°C in a water bath and hold at this temperature for one hour. Shake intermittently
  •  Allow to cool at room temperature
  •  Add 84.0 ml of methanol
  •  Allow it to stand at room temperature for 2 days and shake regularly
  •  Add 0.2 G of Azur II to every 100 ml prepared
  •  Stand for an additional 2 days at room temperature, shake regularly
  •  Filter and store in a dark place.

 Procedure

  • Fix the blood film in absolute methyl alcohol for 2 minutes. This can be done by dipping the smear briefly in a coplin jar containing methyl alcohol.
  • Let the smear to dry.
  • Stain in a 1 in 10 dilution of Giemsa’s stain for 20 minutes. (Add 1 ml of stock Giemsa’s to 9 ml of neutral buffered water)
  • Wash the smear , air dry it and examine under oil immersion objective for detecting the parasite.
Last modified: Monday, 4 June 2012, 5:40 AM