Giemsa's staining
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GIEMSA'S STAINING TECHNIQUE
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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.
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Last modified: Monday, 4 June 2012, 5:40 AM