Hardy Diagnostics Sudan Black B Stain is recommended as a fat stain for bacteria in Burdon's technique, or as a fat stain for animal cells, chromosomes, leukocyte granules, and Golgi apparatus.
Sudan Black B Stain was originally prepared in Germany in the early 1930's. Lison and Dagnelie proposed its use as a myelin stain in 1935. Gerard, also in 1935, first developed its use as a fat stain. In 1940's, both Hartman and Burdon further established its value as a bacterial fat stain. It has also been proposed for staining Golgi apparatus and leukocyte granules.
Sudan Black B Stain is a qualitative method for detecting the presence of fat granules in Bacillus species and other bacteria. This procedure, when performed carefully and consistently, aids in the detection of poly-beta-hydroxybutyrate granules in bacteria.
This product is not intended for primary isolation of patient specimens. It should be used only with cultures of isolated organism. This product is used in conjunction with biochemical tests to identify cultures of isolated organism.
Dry and heat fix the smear to be examined. Shake the Sudan Black B Stain well. Flood the slide with the Sudan Black B, and stain for ten minutes. Drain and blot dry. Wash and clear the smear with xylene or xylene substitute (i.e. Clearene, Cat. no. HDCE016). Counterstain with aqueous safranin for 10-15 seconds. Rinse in tap water and blot dry. Examine under oil immersion for the presence of fat granules. For Legionella pneumophila, counterstain with safranin for one minute.
INTERPRETATION OF RESULTS
When observed microscopically using oil immersion, the cells stain red; the poly-beta-hydroxybutyrate granules, which are highly refractile, stain blue-black in color.
Cat. no. HDZ89