Technique METACHROMATIC GRANULES
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Principle For metachromatic granules (volutin granules) make a smear from a culture grown on Loeffler serum
Method See Neisser's Stain
See Albert's Stain
Positive control  
Negative control  

 

Technique METACHROMATIC GRANULES Neisser's Stain
Principle  
Cautions  
Method Stain with Neisser's stain for 10 seconds.
Rinse rapidly with water.
Stain with 0.2% Bismarck brown or
0.4% chrysoidin for 30 seconds.
Wash rapidly in water, drain or blot dry.
Results Cytoplasm appears light brown. Granules blue-black.
Positive control  
Negative control  
Reagents Neissers stain solution
For use, mix 20 ml of solution A and 10 ml of solution B

Neisser’sSolution A:
Methylene blue 0.1 gm;
95% ethanol 5 ml;
glacial acetic acid 5 ml
distilled water 100 ml.

Dissolve the dye in the water and add the acid and ethanol

Neisser’s Solution B:
Crystal violet 0.33 gm;
95% ethanol 3.3 ml
distilled water 100 ml.

Dissolve the dye in the ethanol- water mixture
Reference  

 

Technique METACHROMATIC GRANULES Albert's Stain
Principle  
Cautions  
Method 1: Stain with Albert's stain fo 3-5 minutes.
2: Rinse with water
3: Blot dry.
3: Stain with Lugol's iodine solution for 1 minute.
4: Rinse with water
5: Drain or blot to dry.
Results Cytoplasm appears light green, granules blue-black
Positive control  
Negative control  
Reagents Albert’s Stain Solution:
Malachite green 0.2 gm;
Toluidine Blue 0.15 gm;
95% ethanol 2 ml;
glacial acetic acid 1 ml;
distilled water 100 ml.

Dissolve the dyes in the ethanol. Mix the acid with the water and add to the dye solution. Let stand for 24 hours and then filter.

Lugol’s Iodine Solution
Reference H. Albert 1921 J. Am. Med Ass 76:240